BR112017011126B1 - COMPOUND, USE THEREOF, AND PHARMACEUTICAL COMPOSITION - Google Patents

Abstract

BILE ACID DERIVATIVES AS FXR/TGR5 AGONISTS AND METHODS OF THEIR USE. The present invention provides amino-substituted bile acid derivative compounds, pharmaceutical compositions comprising such compounds, and methods of using such compounds to prevent or treat FXR-mediated or TGR5-mediated diseases or conditions. In one embodiment, the present invention provides a pharmaceutical composition comprising a therapeutically effective amount of a compound or combination of compounds of the present invention or a pharmaceutically acceptable salt, stereoisomer, solvate, hydrate form or combination thereof, in combination with a pharmaceutically acceptable carrier or excipient.

Description

FIELD OF TECHNIQUE

[0001] The present invention generally relates to compounds and pharmaceutical compositions useful as FXR/TGR5 modulators. Specifically, the present invention relates to bile acid derivatives and methods for their preparation and use.

FUNDAMENTALS OF THE INVENTION

[0002] The Farnesoid X receptor (FXR) is an orphan nuclear receptor initially identified from a rat liver cDNA library (BM. Forman, et al., Cell, 1995, 81(5), 687-693) that is most closely related to the insect ecdysone receptor. FXR is a member of the nuclear receptor family of ligand-activated transcription factors which includes receptors for steroid, retinoid and thyroid hormones ( DJ. Mangelsdorf, et al., Cell, 1995, 83(6), 841-850 ). Relevant physiological ligands of FXR are bile acids (D. Parks et al., Science, 1999, 284(5418), 1362-1365). The most potent is chenodeoxycholic acid (CDCA), which regulates the expression of several genes involved in bile acid homeostasis. Farnesol and derivatives, together called farnesoids, are originally described to activate the mouse ortholog at high concentration but do not activate the human or mouse receptor. FXR is expressed in the liver, throughout the gastrointestinal tract including the esophagus, stomach, duodenum, small intestine, colon, ovary, adrenal gland and kidney. In addition to controlling intracellular gene expression, FXR appears to be also involved in paracrine and endocrine signaling, increasing the expression of cytokine fibroblast growth factor ( J. Holt et al., Genes Dev., 2003, 17(13), 1581-1591 ; T. Inagaki et al., Cell Metab., 2005, 2(4), 217-225 ).

[0003] Small molecule compounds that act as FXR modulators have been disclosed in the following publications: WO 2000/037077, WO 2003/015771, WO 2004/048349, WO 2007/076260, WO 2007/092751, WO 2007/140174, WO 2007/1401 83, WO 2008/051942, WO 2008/157270, WO 2009/005998, WO 2009/012125, WO 2008/025539, WO 2008/025540, WO 2011/020615 and WO 2013/007387.

[0004] Other small molecule FXR modulators have recently been reviewed ( R.C. Buijsman et al. Curr. Med. Chem. 2005, 12, 1017-1075 ).

[0005] The TGR5 receptor is a G protein-coupled receptor that has been identified as a cell surface receptor that responds to bile acids (BAs). The primary structure of TGR5 and its responsiveness to bile acids has been highly conserved in TGR5 among humans, cattle, rabbits, rats and mice and thus suggests that TGR5 has important physiological functions. TGR5 was found to be widely distributed not only in lymphoid tissues but also in other tissues. Elevated levels of TGR5 mRNA were detected in placenta, spleen and monocytes/macrophages. Bile acids have been shown to induce internalization of the TGR5 fusion protein from the cell membrane to the cytoplasm ( Kawamata et al., J. Bio. Chem., 2003, 278, 9435 ). TGR5 was found to be identical to hGPCR19 reported by Takeda et al., FEBS Lett. 2002,520, 97-101.

[0006] TGR5 is associated with the intracellular accumulation of cAMP, which is widely expressed in many cell types. Although activation of this membrane receptor on macrophages decreases production of pro-inflammatory cytokines, (Kawamata, Y., et al., J. Biol. Chem. 2003, 278, 9435-9440) stimulation of TGR5 by BAs in adipocytes and myocytes increases energy expenditure (Watanabe, M., et al. Nature. 2006, 439, 48448 9). This latter effect involves cAMP-dependent induction of iodothyronine deiodinase type 2 (D2), which, locally converting T4 to T3, gives rise to increased thyroid hormone activity. In line with the role of TGR5 in controlling energy metabolism, female TGR5 knockout mice show significant fat accumulation with body weight gain when challenged with a high-fat diet, indicating that a lack of TGR5 decreases energy expenditure and causes obesity (Maruyama, T., et al., J. Endocrinol. 2006, 191, 197-205). Additionally and in line with the involvement of TGR5 in energy homeostasis, bile acid activation of the membrane receptor has been reported to also promote the production of glucagon-like peptide 1 (GLP-1) in murine enteroendocrine cell lines (Katsuma, S., Biochem. Biophys. Res. Commun., 2005, 329, 386-390). Based on all the above observations, TGR5 is an attractive target for the treatment of diseases, for example, obesity, diabetes and metabolic syndrome.

[0007] In addition to the use of TGR5 agonists for the treatment and prevention of metabolic diseases, compounds that modulate TGR5 modulators are also useful for the treatment of other diseases, for example, central nervous system diseases as well as inflammatory diseases (WO 01/77325 and WO 02/84286). TGR5 modulators also provide methods of regulating bile acid and cholesterol homeostasis, fatty acid absorption, and protein and carbohydrate digestion.

[0008] There is a need for the development of FXR and/or TGR5 modulators for the treatment and prevention of diseases. The present invention has identified compounds containing amino, urea, sulfonurea or sulfonamide moieties that modify FXR and/or TGR, as well as methods of using these compounds to treat disease.

SUMMARY OF THE INVENTION

[0009] In one aspect, the invention provides compounds represented by Formula I, or pharmaceutically acceptable salts, stereoisomers, solvates, hydrates or combinations thereof: wherein: Ra is hydrogen or substituted or unsubstituted -C1-C8-alkyl; preferably, Ra is hydrogen or methyl; more preferably, Ra is hydrogen; Rb is selected from the group consisting of 1) Hydrogen; 2) -C(O)NRioRii; 3) -C(O)NHSO2Ri; and 4) -SO2Ri; 5) is selected from the group consisting of: 1) Halogen; 2) Hydroxyl; 3) Substituted or unsubstituted C1 -C8 alkyl; 4) substituted or unsubstituted -C2-C8 alkenyl; 5) Substituted or unsubstituted -C2-C8-alkynyl; 6) substituted or unsubstituted -C3-C8-cycloalkyl; 7) Aryl substituted or unsubstituted; 8) Substituted or unsubstituted arylalkyl; 9) Substituted or unsubstituted heterocycloalkyl; 10) Substituted or unsubstituted heteroaryl; 11) Substituted or unsubstituted heteroarylalkyl; and 12) -NRioRii; 13) is selected from the group consisting of: 1) Hydrogen; 2) Substituted or unsubstituted C1 -C8 alkyl; 3) substituted or unsubstituted -C2-C8 alkenyl; 4) Substituted or unsubstituted -C2-C8-alkynyl; 5) Substituted or unsubstituted arylalkyl; and 6) Substituted or unsubstituted aryl; preferably R2 is hydrogen or methyl. m is selected from 0, 1, 2 and 3, preferably m is 0, 1 or 2. R3 is hydrogen, hydroxyl, -OSO3H, -OSO3-, -OAc, -OPO3H2 or -OPO32; preferably, R3 is hydrogen or hydroxyl. R4 is hydrogen, halogen, CN, N3, hydroxyl, -OSO3H, -OSO3-, -OAc, -OPO3H2, -OPO32-, -SR2 or -NHR2, wherein, R2 is as previously defined; preferably, R4 is hydrogen. Or R3 and R4 are taken together with the carbon atoms to which they are attached to form -CH = CH- or cycloalkyl ring or heterocycloalkyl ring such as, but not limited to, cyclopropyl or epoxide. R5 and R6 are independently selected from hydrogen or a hydroxyl protecting group, such as, but not limited to, acetyl, trimethylsilyl, or benzyl; preferably, R5 and R6 are hydrogen. R7 is selected from the group consisting of: 1) Hydrogen; 2) Halogen; 3) Substituted or unsubstituted -C1-C8-alkyl; 4) substituted or unsubstituted -C2-C8 alkenyl; 5) Substituted or unsubstituted -C2-C8-alkynyl; and 6) substituted or unsubstituted -C3-C8-cycloalkyl; preferably R7 is C1-C4-alkyl, more preferably R7 is ethyl; R10 and R11 are each independently selected from hydrogen, substituted or unsubstituted -C1-C8 alkyl, substituted or unsubstituted -C2 -C8 alkenyl, substituted or unsubstituted -C2 -C8 alkynyl, substituted or unsubstituted -C3 -C8 cycloalkyl, substituted or unsubstituted heterocycloalkyl, or R10 and R11 are taken together with the nitrogen to which they are attached, form a heterocyclic ring; preferably, R11 is hydrogen.

[0010] In another embodiment, the present invention provides a pharmaceutical composition comprising a therapeutically effective amount of a compound or combination of compounds of the present invention or a pharmaceutically acceptable salt form, stereoisomer, solvate, hydrate or combination thereof, in combination with a pharmaceutically acceptable carrier or excipient.

[0011] In another embodiment, the present invention provides a method for preventing or treating an FXR-mediated disease or condition. The method comprises administering a therapeutically effective amount of a compound of the invention. The present invention also provides the use of a compound of the invention for the preparation of a medicament for the prevention or treatment of an FXR-mediated disease or condition.

[0012] In yet another embodiment, the present invention provides a method for preventing or treating a TGR5-mediated disease or condition. The method comprises administering a therapeutically effective amount of a compound of the invention. The present invention also provides the use of a compound of the invention for the preparation of a medicament for preventing or treating a TGR5-mediated disease or condition.

[0013] In certain embodiments, a disease involving TGR5 receptor modulation is selected from metabolic disease, inflammatory disease, liver disease, autoimmune disease, heart disease, kidney disease, cancer, and gastrointestinal disease.

DETAILED DESCRIPTION OF THE INVENTION

[0014] A first embodiment of the invention is a compound represented by Formula I, as described above, or a pharmaceutically acceptable salt, hydrate, solvate, ester or prodrug thereof. In preferred compounds of Formula I, R2, R3, R4, R5, and R6 are hydrogen and R7 is ethyl.

[0015] In another embodiment, the invention provides compounds of Formula I', or pharmaceutically acceptable salts or prodrugs, wherein: Ra is hydrogen; Rb is selected from the group consisting of 1) Hydrogen; 2) -C(O)NRioRii; 3) -C(O)NHSO2Ri; and 4) -SO2Ri; 5) is selected from the group consisting of: 1) Halogen; 2) Hydroxyl; 3) Substituted or unsubstituted C1 -C8 alkyl; 4) substituted or unsubstituted -C2-C8 alkenyl; 5) Substituted or unsubstituted -C2-C8-alkynyl; 6) substituted or unsubstituted -C3-C8-cycloalkyl; 7) Aryl substituted or unsubstituted; 8) Substituted or unsubstituted alkylaryl; 9) Substituted or unsubstituted heterocycloalkyl; 10) Substituted or unsubstituted heteroaryl; 11) Substituted or unsubstituted heteroaryl alkyl; and 12) -NRioRii; 13) is selected from the group consisting of: 1) Hydrogen; 2) Substituted or unsubstituted -C1-C8-alkyl; 3) substituted or unsubstituted -C2-C8 alkenyl; 4) Substituted or unsubstituted -C2-C8-alkynyl; 5) Substituted or unsubstituted alkylaryl; and 6) Substituted or unsubstituted aryl; Preferably R2 is hydrogen or methyl; m is selected from 0, 1, 2 and 3, preferably m is 0, 1 or 2. R3 is hydrogen, hydroxyl, -OSO3H, -OSO3-, -OAc, -OPO3H2 or -OPO32; preferably, R3 is hydrogen or hydroxyl. R4 is hydrogen, halogen, CN, N3, hydroxyl, -OSO3H, -OSO3-, -OAc, -OPO3H2, -OPO32-, -SR2 or -NHR2, wherein, R2 is as previously defined; preferably, R4 is hydrogen. Or R3 and R4 are taken together with the carbon atoms to which they are attached to form -CH = CH- or cycloalkyl ring or heterocycloalkyl ring such as, but not limited to, cyclopropyl or epoxide. 7) and R6 are independently selected from hydrogen or a hydroxyl protecting group, such as, but not limited to, acetyl, trimethylsilyl, or benzyl; preferably, R5 and R6 are hydrogen. 8) is selected from the group consisting of: 1) Hydrogen; 2) Halogen; 3) Substituted or unsubstituted -C1-C8-alkyl; 4) substituted or unsubstituted -C2-C8 alkenyl; 5) Substituted or unsubstituted -C2-C8-alkynyl; and 6) substituted or unsubstituted -C3-C8-cycloalkyl; preferably R7 is C1-C4-alkyl, more preferably R7 is ethyl; R10 and R11 are each independently selected from hydrogen, substituted or unsubstituted -C1-C8 alkyl, substituted or unsubstituted -C2 -C8 alkenyl, substituted or unsubstituted -C2 -C8 alkynyl, substituted or unsubstituted -C3 -C8 cycloalkyl, or R10 and R11 taken together with the nitrogen to which they are attached form a heterocyclic ring; preferably, R11 is hydrogen.

[0016] In preferred embodiments, the compounds of the invention have the stereochemistry set forth in Formula IA: where m, Ra, Rb, R2, R3, R4, R5, R6, and R7 have the meanings given for those variables in Formula I or I'. In certain embodiments, the invention provides a compound represented by Formula II or a pharmaceutically acceptable salt, hydrate, solvate, ester or prodrug thereof: wherein Ra, Rb, R2, R3, R4, R7 and m are as previously defined in Formula I or I'. In certain embodiments, the invention provides a compound represented by Formula III or a pharmaceutically acceptable salt, hydrate, solvate, ester or prodrug thereof: wherein Ra, Rb, R-2, R3, R7 and are as defined above in Formula I or I'.

[0017] Illustrative structures of Formula (III) can be represented, but not limited, by the formula (III-1 ~ III-54), where R1, R7, R10 and are as previously defined in Formula I or I':

[0018] In certain embodiments, the invention provides a compound represented by Formula IV-A, IV-B, IV-C or IV-D or a pharmaceutically acceptable salt, solvate, hydrate, ester or prodrug thereof, wherein R1 and m are as previously defined in Formula I or I'.

[0019] In certain embodiments of the compounds of the invention, R1 is C1-C4-halogenated alkyl, C1-C4-alkyl, C1-C4-alkenyl, substituted or unsubstituted phenyl-C1-C4-alkyl, C3-C6-cycloalkyl, C1-C6-cycloalkyl-C1-C4-alkyl, 5- or 6-membered heterocycloalkyl, substituted or unsubstituted amino, phenyl or halogen.

[0020] In certain embodiments of the compounds of the invention, R1 is ethyl, butyl, t-butyl, propyl, benzyl, vinyl, allyl, or fluoro; or Ri is methyl, isopropyl or phenyl. In certain embodiments of the compounds of the invention, R1 is dimethylamino or p-tert-butylphenyl.

[0021] In certain embodiments of the compounds of the invention, R1 is selected from the groups defined in the table below:

[0022] In certain embodiments of the compounds of the invention, R11 is hydrogen and R10 is hydrogen, C1-C4 alkyl, halogenated C1-C4 alkyl, C1-C4 alkenyl, substituted or unsubstituted phenyl-C1-C4-alkyl, C3-C6-cycloalkyl, C1-C6-cycloalkyl-C1-C4-alkyl, 5- or 6-membered heterocycloalkyl, or substituted or unsubstituted phenyl.

[0023] In certain embodiments of the compounds of the invention, R11 is hydrogen and R10 is hydrogen, methyl, ethyl, isopropyl, butyl, t-butyl, propyl, benzyl, vinyl, allyl,

[0024] Representative compounds of the invention include, but are not limited to, the following compounds (compounds 1 to 75 in Table 1), according to Formula IV-A, wherein R1 and are outlined for each compound in Table 1. Table 1

[0025] Representative compounds of the invention include, but are not limited to, the following compounds (compounds 76 to 150 in Table 2), according to Formula IV-B, wherein R1 and m are outlined for each compound in Table 2. Table 2

[0026] In certain embodiments, the invention provides a compound represented by Formula VA and VB or a pharmaceutically acceptable salt, solvate, hydrate, ester or prodrug thereof. wherein R1 and m are as previously defined in Formula I or I'.

[0027] Representative compounds of the invention include, but are not limited to, the following compounds (compounds 151 to 225 in Table 3), according to Formula VA, wherein R1 and m are outlined for each compound in Table 3. Table 3

[0028] Representative compounds of the invention include, but are not limited to, the following compounds (compounds 226 to 300 in Table 4), according to Formula VB, wherein R1 and m are outlined for each compound in Table 4. Table 4

[0029] In certain embodiments, the invention provides a compound represented by Formula VI-A or VI-B or a pharmaceutically acceptable salt, solvate, hydrate, ester or prodrug thereof: where R10 and m are as defined above. In Formula I or I'

[0030] Representative compounds of the invention include, but are not limited to, the following compounds (compounds 301 to 375 in Table 5), according to Formula VI-A, wherein R10 and m are outlined for each compound in Table 5. Table 5

[0031] Representative compounds of the invention include, but are not limited to, the following compounds (compounds 376 to 450 in Table 6), according to Formula VI-B, wherein R10 and m are outlined for each compound in Table 6. Table 6

[0032] Representative compounds of the invention include, but are not limited to, the following compounds (compounds 451 to 525 in Table 7), according to Formula IV-C, wherein R1 and m are outlined for each compound in Table 7. Table 7

[0033] Representative compounds of the invention include, but are not limited to, the following compounds (compounds 526 to 600 in Table 8), according to Formula IV-D, wherein R1 and m are outlined for each compound in Table 8. Table 8

[0034] In certain embodiments, the invention provides a compound represented by Formula VII-A or VII-B or a pharmaceutically acceptable salt, solvate, hydrate, ester or prodrug thereof: wherein R1 and m are as previously defined in Formula I or I'.

[0035] Representative compounds of the invention include, but are not limited to, the following compounds (compounds 601 to 675 in Table 9), according to Formula VII-A, wherein R1 and m are outlined for each compound in Table 9. Table 9

[0036] Representative compounds of the invention include, but are not limited to, the following compounds (compounds 676 to 750 in Table 10), according to Formula VII-A, wherein R1 and m are outlined for each compound in Table 10.

[0037] In certain embodiments, the present invention provides a method for preventing or treating an FXR-mediated disease or condition. The method comprises administering a therapeutically effective amount of a compound of the invention. The present invention also provides the use of a compound of the invention for the preparation of a medicament for the prevention or treatment of an FXR-mediated disease or condition.

[0038] In certain embodiments, the FXR-mediated disease or condition is cardiovascular disease, atherosclerosis, arteriosclerosis, hypercholesteremia or hyperlipidemia, chronic liver disease, gastrointestinal disease, kidney disease, metabolic disease, cancer (i.e., colorectal cancer) or neurological indications such as stroke.

[0039] In certain embodiments, the chronic liver disease is primary biliary cirrhosis (PBC), cerebrotendenous xanthomatosis (CTX), primary sclerosing cholangitis (PSC), drug-induced cholestasis, intrahepatic cholestasis of pregnancy, cholestasis associated with parenteral nutrition (PNAC), bacterial overgrowth or cholestasis associated with sepsis, autoimmune hepatitis, chronic viral hepatitis, alcoholic liver disease, non-alcoholic fatty liver disease (NAFLD), nonalcoholic steatohepatitis (NASH), liver transplant graft-associated host disease, live-donor transplant liver regeneration, congenial liver fibrosis, liver gallstones, intra- or extrahepatic malignancy, Sjogren's syndrome, sarcoidosis, Wilson's disease, Gaucher's disease, hemochromatosis, or alpha-1-antitrypsin deficiency. In certain embodiments, the gastrointestinal disease is inflammatory bowel disease (IBD) (including Crohn's disease and ulcerative colitis), irritable bowel syndrome (IBS), bacterial overgrowth, malabsorption, post-radiation colitis, or microscopic colitis.

[0040] In some embodiments, the kidney disease is diabetic nephropathy, focal segmental glomerulosclerosis (FSGS), hypertensive nephrosclerosis, chronic glomerulonephritis, chronic transplant glomerulopathy, chronic interstitial nephritis, or polycystic kidney disease.

[0041] In certain embodiments, the cardiovascular disease is atherosclerosis, arteriosclerosis, dyslipidemia, hypercholesterolemia, or hypertriglyceridemia.

[0042] In certain embodiments, the metabolic disease is insulin resistance, Type I and Type II diabetes or obesity.

[0043] In yet another embodiment, the invention provides the use of the compound or pharmaceutical composition of the invention in the manufacture of a medicament for treating or preventing a disease in a subject that involves TGR5 receptor modulation. The invention includes a method of treating or preventing a disease that involves modulating the TGR5 receptor in a subject by administering a compound or pharmaceutical composition of the invention.

[0044] In certain embodiments, a disease involving TGR5 receptor modulation is selected from metabolic disease, inflammatory disease, liver disease, autoimmune disease, heart disease, kidney disease, cancer, and gastrointestinal disease.

[0045] In one aspect, the invention provides the use, wherein the disease is an inflammatory disease selected from allergy, osteoarthritis, appendicitis, bronchial asthma, pancreatitis, allergic rash and psoriasis. The invention includes a method of treating or preventing an inflammatory disease selected from allergy, osteoarthritis, appendicitis, bronchial asthma, pancreatitis, allergic rash and psoriasis.

[0046] In one aspect, the invention provides the use, wherein the disease is an autoimmune disease selected from rheumatoid arthritis, multiple sclerosis and type I diabetes. The invention includes a method of treating or preventing an autoimmune disease selected from rheumatoid arthritis, multiple sclerosis and type I diabetes.

[0047] In one aspect, the invention provides the use, wherein the disease is a gastrointestinal disease selected from inflammatory bowel disease (Crohn's disease, ulcerative colitis), short bowel syndrome (post-radiation colitis), microscopic colitis, irritable bowel syndrome (malabsorption) and bacterial overgrowth. The invention includes a method of treating or preventing a gastrointestinal disease selected from inflammatory bowel disease (Crohn's disease, ulcerative colitis), short bowel syndrome (post-radiation colitis), microscopic colitis, irritable bowel syndrome (malabsorption) and bacterial overgrowth.

[0048] In one aspect, the invention provides the use, wherein the disease is kidney disease selected from diabetic nephropathy, chronic renal failure, hypertensive nephrosclerosis, chronic glomerulonephritis, chronic transplant glomerulopathy, chronic interstitial nephritis and polycystic kidney disease. The invention includes a method for treating or preventing kidney disease selected from diabetic nephropathy, chronic renal failure, hypertensive nephrosclerosis, chronic glomerulonephritis, chronic transplant glomerulopathy, chronic interstitial nephritis and polycystic kidney disease.

[0049] In one aspect, the invention provides the use, wherein the disease is cancer selected from colorectal cancer, liver cancer, hepatocellular carcinoma, cholangio carcinoma, kidney cancer, gastric cancer, pancreatic cancer, prostate cancer and insulanoma. The invention includes a method of treating or preventing cancer selected from colorectal cancer, liver cancer, hepatocellular carcinoma, cholangio carcinoma, kidney cancer, gastric cancer, pancreatic cancer, prostate cancer and insulanoma.

[0050] In one aspect, the compound is a selective FXR agonist over TGR5 activator.

[0051] In one aspect, the compound is a selective TGR5 agonist over FXR activator.

[0052] In one aspect, the compound is a dual agonist for FXR and TGR5.

[0053] Yet a further aspect of the present invention is a process of making any of the compounds outlined herein employing any of the synthetic means outlined herein.

DEFINITIONS

[0054] Listed below are definitions of various terms used to describe this invention. These definitions apply to the terms as they are used throughout this specification and claims, unless otherwise limited in specific cases, both individually and as part of a larger group.

[0055] The term "alkyl", as used herein, refers to a straight- or branched-chain, monovalent, saturated hydrocarbon group. Preferred alkyl radicals include C1-C6 alkyl and C1-C8 alkyl radicals. Examples of C1-C6 alkyl groups include, but are not limited to, methyl, ethyl, propyl, isopropyl, n-butyl, tert-butyl, neopentyl, n-hexyl; and examples of C1C8 alkyl include, but are not limited to, methyl, ethyl, propyl, isopropyl, n-butyl, tert-butyl, neopentyl, n-hexyl, heptyl, and octyl.

[0056] The term "alkenyl", as used herein, designates a monovalent group derived from a hydrocarbon moiety by the removal of a single hydrogen atom in which the hydrocarbon moiety has at least one carbon-carbon double bond. Preferred alkenyl groups include C2-C6 alkenyl and C2-C8 alkenyl groups. Alkenyl groups include, among others, for example, ethenyl, propenyl, butenyl, 1-methyl-2-buten-1-yl, heptenyl, octenyl and the like.

[0057] The term "alkynyl", as used herein, designates a monovalent group derived from a hydrocarbon moiety by the removal of a single hydrogen atom in which the hydrocarbon moiety has at least one carbon-carbon triple bond. Preferred alkynyl groups include C2-C6 alkynyl and C2-C8 alkynyl groups. Representative alkynyl groups include, but are not limited to, for example, ethynyl, 1-propynyl, 1-butynyl, heptynyl, octynyl, and the like.

[0058] The term "carbocycle" refers to a saturated (e.g., "cycloalkyl"), partially saturated (e.g., "cycloalkenyl" or "cycloalkynyl"), or completely unsaturated (e.g., "aryl") ring system containing zero heteroatom ring atoms. "Ring atoms" or "ring members" are those atoms bonded together to form a ring or rings. When a carbocycle group is a divalent moiety linking two other elements in a depicted chemical structure (such as Z in Formula IA), the carbocycle group may be linked to the other two elements through any two substitutable ring atoms. AC4-C6 carbocycle has 4-6 ring atoms.

[0059] The term "cyclic alkyl" used herein denotes a monovalent group derived from a saturated monocyclic or polycyclic carbocyclic ring compound by the removal of a single hydrogen atom. Preferred cycloalkyl groups include C3-C8 cycloalkyl groups and C3-C12 cycloalkyl groups. Examples of C3-C8-cycloalkyl include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclopentyl and cyclooctyl; and examples of C3-C12cycloalkyl include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, bicyclo[2.2.1]heptyl and bicyclo[2.2.2]octyl.

[0060] The term "cycloalkenyl", as used herein, denotes a monovalent group derived from a carbocyclic, monocyclic or polycyclic ring compound that possesses at least one carbon-carbon double bond by the removal of a single hydrogen atom. Preferred cycloalkenyl groups include C3-C8 cycloalkenyl and C3-C12 cycloalkenyl groups. Examples of C3-C8 cycloalkenyl include, but are not limited to, cyclopropenyl, cyclobutenyl, cyclopentenyl, cyclohexenyl, cycloheptenyl, cyclooctenyl and the like; and examples of C3-C12 cycloalkenyl include, but are not limited to, cyclopropenyl, cyclobutenyl, cyclopentenyl, cyclohexenyl, cycloheptenyl, cyclooctenyl and the like.

[0061] The term "aryl" used herein refers to a mono- or bicyclic carbocyclic ring system having one or more aromatic rings, including, but limited to, phenyl, naphthyl, tetrahydronaphthyl, indanyl, indenyl and the like.

[0062] The term "arylalkyl", as used herein, refers to a C1-C3 alkyl or C1-C6 alkyl residue coupled to an aryl ring. Examples include, but are not limited to, benzyl, phenethyl and the like.

[0063] The term "heteroaryl", as used herein, refers to a mono-, bi- or tricyclic aromatic radical or ring having between five to ten ring atoms, of which at least one ring atom is selected from S, O and N; wherein any N or S contained within the ring can optionally be oxidized. Preferred heteroaryl groups are monocyclic or bicyclic. Heteroaryl groups include, but are not limited to, pyridinyl, pyrazinyl, pyrimidinyl, pyrrolyl, pyrazolyl, imidazolyl, thiazolyl, oxazolyl, iso-oxazolyl, thiadiazolyl, oxadiazolyl, thiophenyl, furanyl, quinolinyl, isoquinolinyl, benzimidazolyl, benzoxazolyl, quinoxalinyl, and the like.

[0064] The term "heteroarylalkyl", as used herein, refers to a C1-C3 alkyl or C1-C6 alkyl residue coupled to a heteroaryl ring. Examples include, but are not limited to, pyridinylmethyl, pyrimidinylethyl and the like.

[0065] The term "substituted" as used herein, refers to the independent replacement of one, two, three or more hydrogen atoms present with substituents including, but not limited to, deuterium, -F, -Cl, -Br, -I, -OH, protected hydroxy, -NO2, -CN, -NH2, N3, protected amino, alkoxy, thioalkoxy, oxo, -halo-C1-C12-alkyl, -halo-C2- -C12-alkenyl, -halo-C2-C12-alkynyl, -halo-C3-C12-cycloalkyl, -NH -C1-C12-alkyl, -NH -C2-C12-alkenyl, -NH -C2-C12-alkynyl, -NH -C3-C12-cycloalkyl, -NH -aryl, -NH -heteroaryl, -NH -heterocycloalkyl, -dialkylamino, -diarylamino, -diheteroarylamino, -O-C1-C12-alkyl, -O-C2-C12-alkenyl, -O-C2-C12-alkynyl, -O-C3-C12-cycloalkyl, -O-aryl, -O-heteroaryl, -O-heterocycloalkyl, -C(O)-C1-C12-alkyl, -C(O)-C2-C12-alkyl nyl, -C(O)-C2-C12-alkynyl, -C(O)-C3-C12-cycloalkyl, -C(O)-aryl, -C(O)-heteroaryl, -C(O)-heterocycloalkyl, -CONH2, -CONH-C1-C12-alkyl, -CONH-C2-C12-alkenyl, -CONH-C2-C12-alkynyl, -CONH-C 3-C12-cycloalkyl, -CONH-aryl, -CONH-heteroaryl, -CONH-heterocycloalkyl, -OCO2-C1-C12-alkyl, -OCO2-C2-C12-alkenyl, -OCO2-C2-C12-alkynyl, -OCO2-C3-C12-cycloalkyl, -OCO2-aryl, -OCO2-heteroaryl, -OCO2- heterocycloalkyl, -OCONH2, -OCONH-C1-C12-alkyl, -OCONH-C2-C12-alkenyl, -OCONH-C2-C12-alkynyl, -OCONH-C3-C12-cycloalkyl, -OCONH-aryl, -OCONH-heteroaryl, -OCONH-heterocycloalkyl, -NHC(O)-C1-C12-alkyl, -NHC (O)-C2-C12-alkenyl, -NHC(O)-C2-C12-alkynyl, -NHC(O)-C3-C12-cycloalkyl, -NHC(O)-aryl, -NHC(O)-heteroaryl, -NHC(O)-heterocycloalkyl, -NHCO2-C1-C12-alkyl, -NHCO2-C2-C12-alkenyl, -NHCO2- C2-C12-alkynyl, -NHCO2-C3-C12-cycloalkyl, -NHCO2-aryl, -NHCO2-heteroaryl, -NHCO2-heterocycloalkyl, -NHC(O)NH2, -NHC(O)NH-C1-C12-alkyl, -NHC(O)NH-C2-C12-alkenyl, -NHC(O)NH-C2-C12-alkynyl, -NHC (O)NH-C3-C12-cycloalkyl, -NHC(O)NH-aryl, -NHC(O)NH- heteroaryl, -NHC(O)NH-heterocycloalkyl, NHC(S)NH2, -NHC(S)NH-C1-C12-alkyl, -NHC(S)NH-C2-C12-alkenyl, -NHC(S)NH-C2-C12-alkynyl, -NHC(S) )NH-C3-C12-cycloalkyl, -NHC(S)NH-aryl, -NHC(S)NH-heteroaryl, -NHC(S)NH-heterocycloalkyl, -NHC(NH)NH2, -NHC(NH)NH-C1-C12-alkyl, -NHC(NH)NH-C2-C12-alkenyl, -NHC(NH)NH-C2-C12-alkynyl, -NHC(NH) -NH-C3-C12-cycloalkyl, -NHC(NH)NH-aryl, -NHC(NH)NH-heteroaryl, -NHC(NH)NH-heterocycloalkyl, -NHC(NH)-C1-C12-alkyl, -NHC(NH)-C2-C12-alkenyl, -NHC(NH)-C2-C12-alkynyl, -NHC(NH)-C3-C12-cycloalkyl, - NHC(NH)-aryl, -NHC(NH)-heteroaryl, -NHC(NH)-heterocycloalkyl, -C(NH)NH-C1-C12-alkyl, -C(NH)NH-C2-C12-alkenyl, -C(NH)NH-C2-C12-alkynyl, -C(NH)NH-C3-C12-cycloalkyl, -C(NH)NH-aryl, -C(NH)NH- heteroaryl, -C(NH)NH-heterocycloalkyl, -S(O)-C1-C12-alkyl, - S(O)-C2-C12-alkenyl, - S(O)-C2-C12-alkynyl, - S(O)-C3-C12-cycloalkyl, - S(O)-aryl, - S(O)-heteroaryl, - S(O)-heterocycloalkyl-SO2NH2, -SO2NH-C1-C12-alkyl, -SO2NH-C2-C12-alkenyl, -SO2NH-C2-C12-alkynyl, -SO2NH-C3-C12-cycloalkyl, -SO2NH-aryl, -SO2NH-heteroaryl, -SO2NH-heterocycloalkyl, -NHSO2-C1-C12-alkyl, -NHSO2-C2-C12-alkenyl, -NHSO2-C2-C12-alkynyl, -NHSO2-C3-C12-cycloalkyl, -NHSO2-aryl, -NHSO2-heteroaryl, -NHSO2-heterocycloalkyl, -CH2NH2, -CH2SO2CH3, -aryl, -arylalkyl, -heteroaryl, -heteroarylalkyl, -heterocycloalkyl, -C3-C12-cycloalkyl, polyalk xyalkyl, polyalkoxy, -methoxymethoxy, -methoxyethoxy, -SH, -S-C1-C12-alkyl, -S-C2-C12-alkenyl, -S-C2-C12-alkynyl, -S-C3-C12-cycloalkyl, -S-aryl, -S-heteroaryl, -S-heterocycloalkyl, methylthiomethyl, or -L'-R', wherein L' is C1- C6 alkylene, C2-C6alkenylene or C2-C6alkynylene, and R' is aryl, heteroaryl, heterocyclic, C3-C12cycloalkyl or C3-C12cycloalkenyl. It is understood that aryl, heteroaryl, alkyl and the like may be additionally substituted. In some cases, each substituent on a substituted moiety is further optionally substituted with one or more groups, with each group independently selected from -F, -Cl, -Br, -I, -OH, -NO2, -CN, or -NH2.

[0066] According to the invention, any aryl, substituted aryl, heteroaryl and substituted heteroaryl described herein may be any substituted aromatic group. Aromatic groups can be substituted or unsubstituted.

[0067] It is understood that any alkyl, alkenyl, alkynyl, cycloalkyl and cycloalkenyl moiety described herein may also be an aliphatic group, an alicyclic group or a heterocyclic group. An "aliphatic group" is a non-aromatic unit which may contain any combination of carbon atoms, hydrogen atoms, halogen atoms, oxygen, nitrogen or other atoms and optionally contains one or more units of unsaturation, e.g. double and/or triple bonds. An aliphatic group can be straight chain, branched, or cyclic and preferably contains from about 1 to about 24 carbon atoms, more typically from about 1 to about 12 carbon atoms. In addition to aliphatic hydrocarbon groups, aliphatic groups include, for example, polyalkoxyalkyl such as polyalkylene glycol, polyamines and polyimines, for example. These aliphatic groups can be substituted below. It is understood that aliphatic groups may be used in place of the alkyl, alkenyl, alkynyl, alkylene, alkenylene and alkynylene groups described herein.

[0068] The term "alicyclic" used herein denotes a monovalent group derived from a monocyclic or polycyclic carbocyclic ring compound saturated by the removal of a single hydrogen atom. Examples include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, bicyclo[2.2.1]heptyl, and bicyclo[2.2.2]°Ctyl. These alicyclic groups can be further substituted.

[0069] The term "heterocycloalkyl" and "heterocyclic" may be used interchangeably and refers to a non-aromatic 3-, 4-, 5-, 6-, or 7-membered ring or a bi- or tricyclic fusion system wherein: (i) each ring contains between one and three heteroatoms independently selected from oxygen, sulfur, and nitrogen, (ii) each 5-membered ring has 0 to 1 double bonds and each ring has 6 members have 0 to 2 double bonds, (iii) the nitrogen and sulfur heteroatoms can be optionally oxidized, (iv) the nitrogen heteroatom can optionally be quaternized, (v) any of the above rings can be fused with a benzene ring, and (vi) the remaining ring atoms are carbon atoms that can optionally be oxo-substituted. Representative heterocyclicalkyl groups include, but are not limited to, [1,3]dioxolane, pyrrolidinyl, pyrazolinyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, piperidinyl, piperazinyl, oxazolidinyl, isoxazolidinyl, morpholinyl, thiazolidinyl, isothiazolidinyl, quinoxalinyl, pyridazinonyl, and tetrahydrofuryl. Such heterocyclic groups can be further substituted to give substituted heterocyclic.

[0070] It will be apparent that, in various embodiments of the invention, substituted or unsubstituted alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, cycloalkynyl, arylalkyl, heteroarylalkyl and heterocycloalkyl may be monovalent or divalent. Thus, the alkylene, alkenylene and alkynylene, cycloalkylene, cycloalkenylene, cycloalkylene, arylalkylene, heteroarylalkylene and heterocycloalkylene groups are to be included in the above definitions and are applicable to provide the formulas herein with proper valency.

[0071] The term "hydroxy activating group" used herein refers to an unstable chemical moiety that is known in the art to activate a hydroxy group so that it will break down during synthetic procedures such as in substitution or elimination reactions. Examples of hydroxy activating group include, but are not limited to, mesylate, tosylate, triflate, p-nitrobenzoate, phosphonate, and the like.

[0072] The term "activated hydroxy" used herein refers to a hydroxy group activated with a hydroxy activating group as defined above, including mesylate, tosylate, triflate, p-nitrobenzoate, phosphonate groups, for example.

[0073] The term "protected hydroxy" used herein refers to a hydroxy group protected with a hydroxy protecting group, defined above, including benzoyl, acetyl, trimethylsilyl, triethylsilyl, methoxymethyl groups.

[0074] The term "hydroxy protecting group" used herein refers to an unstable chemical moiety that is known in the art to protect a hydroxy group against undesirable reactions during synthetic procedures. After said synthetic procedures, the hydroxy protecting group, as described here, can be selectively removed. Protecting groups known in the art are generally described in T.H. Greene and P.G., S.M. Wuts, Protective Groups in Organic Synthesis, 3rd edition, John Wiley & Sons, New York (1999). Examples of hydroxy protecting groups include benzyloxycarbonyl, 4-nitrobenzyloxycarbonyl, 4-bromobenzyloxycarbonyl, 4-methoxybenzyloxycarbonyl, methoxycarbonyl, tert-butoxycarbonyl, isopropoxycarbonyl, diphenylmethoxycarbonyl, 2,2,2-trichloroethoxycarbonyl, 2-(trimethylsilyl)ethoxycarbonyl, 2-furfuryloxycarbonyl, allyloxycarbonyl, acetyl, formyl, chloroacetyl, trifluoroacetyl, methoxyacetyl, phenoxyacetyl, benzoyl, methyl, t-butyl, 2,2,2-trichloroethyl, 2-trimethylsilylethyl, 1,1-dimethyl-2-propenyl, 3-methyl-3-butenyl, allyl, benzyl, para-methoxybenzyldiphenylmethyl, triphenylmethyl (trityl), tetrahydrofuryl, methoxymethyl, methyl thiomethyl, benzyloxymethyl, 2,2,2-trichloroethoxymethyl, 2-(trimethylsilyl)ethoxymethyl, methanesulfonyl, para-toluenesulfonyl, trimethylsilyl, triethylsilyl, triisopropylsilyl and the like. Preferred hydroxy protecting groups for the present invention are acetyl (Ac or -C(O)CH3), benzoyl (Bz or -C(O)C6H5), and trimethylsilyl (TMS or -Si(CH3)3).

[0075] The terms "halogen" or "halo", as used herein, refer to an atom selected from fluorine, chlorine, bromine and iodine.

[0076] The compounds described herein contain one or more asymmetric centers and can therefore give rise to enantiomers, diastereoisomers and other stereoisomeric forms that can be defined, in terms of absolute stereochemistry, as (R)- or (S)- or as (D)- or (L)- for amino acids. The present invention is intended to include all possible isomers, as well as their racemic and optically pure forms. Optical isomers can be prepared from their respective optically active precursors by the procedures described above or by resolving the racemic mixtures. Resolution can be performed in the presence of a resolving agent, by chromatography or by repeated crystallization or by some combination of these techniques which are known to those skilled in the art. More details on resolutions can be found in Jacques, et al., Enantiomers, Racemates, and Resolutions (John Wiley & Sons, 1981). When the compounds described herein contain olefinic double bonds or other centers of geometric asymmetry, and unless otherwise specified, the compounds are intended to include both Z and E geometric isomers. Likewise, all tautomeric forms are also to be included. The configuration of any carbon-carbon double bond appearing here is selected for convenience only and is not intended to designate a particular configuration unless the text indicates otherwise; thus, a carbon-carbon double bond arbitrarily represented here as trans can be cis, trans, or a mixture of the two in any proportion.

[0077] The term "individual" herein refers to a mammal. An individual also refers to, for example, dogs, cats, horses, cows, pigs, guinea pigs and the like. Preferably, the subject is a human. When the subject is a human being, the subject may be referred to herein as a patient.

[0078] As used herein, the term "pharmaceutically acceptable salt" refers to those salts of the compounds, formed by the process of the present invention, which are, within the scope of good medical judgment, suitable for use in contact with the tissues of humans and lower mammals without toxicity, irritation, undue allergic response, and the like, and are commensurate with a reasonable benefit/risk ratio. Pharmaceutically acceptable salts are well known in the art.

[0079] Berge, et al. describe pharmaceutically acceptable salts in detail in J. Pharmaceutical Sciences, 66: 1-19 (1977). Salts can be prepared in situ during isolation and purification of the compounds of the invention or separately by reacting the free base with a suitable organic or inorganic acid. Examples of pharmaceutically acceptable salts include, but are not limited to, non-toxic acid addition salts, for example, are salts of an amino group formed with inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid and perchloric acid or with organic acids such as acetic acid, maleic acid, tartaric acid, citric acid, succinic acid or malonic acid or using other methods used in the art such as ion exchange. Other pharmaceutically acceptable salts include, but are not limited to, adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate, gluconate, hemisulfate, heptanoate salts , hexanoate, hydroiodide, 2-hydroxyethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate, pivalate, propionate , stearate, succinate, sulfate, tartrate, thiocyanate, p-toluenesulfonate, undecanoate, valerate salts and the like. Representative alkali or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium and the like. In addition pharmaceutically acceptable salts include, where appropriate, non-toxic ammonium, quaternary ammonium, and amine cations formed using counterions such as halide, hydroxide, carboxylate, sulfate, phosphate, nitrate, alkyl of 1 to 6 carbon atoms and aryl sulfonate and sulfonate.

[0080] Pharmaceutically acceptable salts can also be prepared by deprotonating the parent compound with a suitable base, thereby forming the anionic conjugate base of the parent compound. In such salts, the counter ion is a cation. Suitable cations include ammonium and metal cations, such as alkali metal cations, including Li+, Na+, K+ and Cs+, and alkaline earth metal cations, such as Mg2+ and Ca2+.

[0081] The term "amino protecting group" used herein refers to an unstable chemical moiety that is known in the art to protect an amino group against undesirable reactions during synthetic procedures. After said synthetic procedures, the amino protecting group, as described here, can be selectively removed. Amino protecting groups known in the art are generally described in T.H. Greene and P.G. M. Wuts, Protective Groups in Organic Synthesis, 3rd edition, John Wiley & Sons, New York (1999). Examples of amino protecting groups include, but are not limited to, butoxycarbonyl, 9-fluorenylmethoxycarbonyl, benzyloxycarbonyl, and the like.

[0082] As used herein, the term "pharmaceutically acceptable ester" refers to esters of the compounds formed by the process of the present invention that hydrolyze in vivo and include those that readily break down in the human body to leave the parent compound or salt thereof. Suitable ester groups include, for example, those derived from pharmaceutically acceptable aliphatic carboxylic acids, particularly alkanoic, alkenoic, cycloalkanoic and alkanedioic acids, in which each alkyl or alkenyl unit advantageously has no more than 6 carbon atoms. Examples of particular esters include but are not limited to formates, acetates, propionates, butyrates, acrylates and ethylsuccinates.

[0083] As used herein, the term "pharmaceutically acceptable prodrugs" refers to those prodrugs of the compounds by the process of the present invention that are, within the scope of good medical judgment, suitable for use in contact with the tissues of humans and smaller animals without inappropriate toxicity, irritation, allergic response and the like, and are commensurate with a reasonable benefit/risk ratio and are effective for the intended use, as well as zwitterionic forms, if possible, of the compounds of the present invention. "Prodrug" as used herein means a compound that is convertible in vivo by metabolic means (e.g., by hydrolysis) to yield any compound encompassed by the formulas of the present invention. Various forms of prodrugs are known in the art, for example, as discussed in Bundgaard, (ed.), Design of Prodrugs, Elsevier (1985); Widder, et al. (ed.), Methods in Enzymology, Vol. 4, Academic Press (1985); Krogsgaard-Larsen, et al., (ed). "Design and Application of Prodrugs, Textbook of Drug Design and Development, Chapter 5, 113-191 (1991); Bundgaard, et al., Journal of Drug Deliver Reviews, 8:1-38 (1992); Bundgaard, J. of Pharmaceutical Sciences, 77:285 et seq. (1988); Higuchi and Stella (eds.) Prodrugs as Novel Drug Delivery Systems, American Chemical Society (1975) and Bernard Testa & Joachim Mayer, “Hydrolysis In Drug And Prodrug Metabolism: Chemistry, Biochemistry And Enzymology,” John Wiley and Sons, Ltd. (2002).

[0084] The term "treatment", as used herein, means alleviating, lessening, reducing, eliminating, modulating or improving, i.e., causing regression of the disease state or condition. Treatment may also include inhibiting, i.e., arresting the development of, an existing disease state or condition and alleviating or ameliorating, i.e., causing regression of an existing disease state or condition, for example, when the disease state or condition is already present.

[0085] The term "prevent" as used herein means completely or nearly completely preventing a disease state or condition from occurring in a patient or individual, especially when the patient or individual is predisposed to such or at risk of contracting a disease state or condition.

[0086] Additionally, the compounds of the present invention, for example, the salts of the compounds, can exist in hydrated or non-hydrated (anhydrous) form or as solvates with other solvent molecules. Non-limiting examples of hydrates include monohydrates, dihydrates, and the like. Non-limiting examples of solvates include ethanol solvates, acetone solvates, and the like.

[0087] "Solvates" means solvent addition forms which contain stoichiometric or non-stoichiometric amounts of solvent. Some compounds have a tendency to capture a fixed molar ratio of solvent molecules in the crystalline solid state, thus forming a solvate. If the solvent is water, the solvate formed is a hydrate, when the solvent is alcohol, the solvate formed is an alcoholate. Hydrates are formed by the combination of one or more water molecules with one of the substances in which water maintains its molecular state as H2O, this combination being capable of forming one or more hydrates.

[0088] As used herein, the term "analogue" refers to a chemical compound that is structurally similar to another but differs slightly in composition (as in the replacement of an atom by an atom of a different element or in the presence of a particular functional group, or the replacement of one functional group by another functional group). Thus, an analogue is a compound that is similar or comparable in function and appearance to the reference compound.

[0089] The term "aprotic solvent" used herein refers to a solvent that is relatively inert to proton activity, ie, does not act as a proton donor. Examples include but are not limited to hydrocarbons such as hexane and toluene, for example, halogenated hydrocarbons such as methylene chloride, ethylene chloride, chloroform and the like, heterocyclic compounds such as tetrahydrofuran and N-methylpyrrolidinone, and ethers such as diethyl ether, bis-methoxymethyl ether. Such solvents are well known to those skilled in the art, and individual solvents or mixtures thereof may be preferred for specific compounds and reaction conditions, depending on factors such as the solubility of the reagents, the reactivity of the reagents, and preferred temperature ranges, for example. Further discussion of aprotic solvents can be found in organic chemistry textbooks or specialized monographs, for example: Organic Solvents Physical Properties and Methods of Purification, 4th ed., edited by John A. Riddick et al., Vol. II, in Techniques of Chemistry Series, John Wiley & Sons, NY, 1986.

[0090] The terms "protogenic organic solvent" or "protic solvent" used herein refer to a solvent that tends to donate protons, such as an alcohol, for example, methanol, ethanol, propanol, isopropanol, butanol, t-butanol, and the like. Such solvents are well known to those skilled in the art, and individual solvents or mixtures thereof may be preferred for specific compounds and reaction conditions, depending on factors such as the solubility of the reagents, the reactivity of the reagents, and preferred temperature ranges, for example. Further discussion of aprotic solvents can be found in organic chemistry textbooks or specialized monographs, for example: Organic Solvents Physical Properties and Methods of Purification, 4th ed., edited by John A. Riddick et al., Vol. II, in Techniques of Chemistry Series, John Wiley & Sons, NY, 1986.

[0091] Combinations of substituents and variables envisioned by this invention are preferably those that result in the formation of stable compounds. The term "stable" used herein refers to compounds that possess sufficient stability to permit manufacture and which maintain the integrity of the compound for a period of time sufficient to be useful for the purposes detailed herein (e.g., therapeutic or prophylactic administration to an individual).

[0092] The synthesized compounds can be separated from a reaction mixture and subsequently are purified by a method such as column chromatography, high pressure liquid chromatography or recrystallization. Furthermore, the various synthetic steps can be carried out in an alternate sequence or order to generate the desired compounds. In addition, solvents, temperatures, reaction times, etc. outlined herein are for illustrative purposes only and varying reaction conditions can produce the desired bridged macrocyclic products of the present invention. Synthetic chemistry transformations and protecting group (protection and deprotection) methodologies useful in synthesizing the compounds described herein include, for example, those described in R. Larock, Comprehensive Organic Transformations, VCH Publishers (1989); T.W. Greene and P.G.M. Wuts, Protective Groups in Organic Synthesis, 2d. Ed., John Wiley and Sons (1991); L. Fieser and M. Fieser, Fieser and Fiesers Reagents for Organic Synthesis, John Wiley and Sons (1994); and L. Paquette, ed., Encyclopedia of Reagents for Organic Synthesis, John Wiley and Sons (1995).

[0093] The compounds of this invention can be modified by adding various functionalities through synthetic means outlined here to enhance selective biological properties. Such modifications include those that increase biological penetration into a given biological system (e.g., blood, lymphatic system, central nervous system), increase oral availability, increase solubility to allow administration by injection, alter metabolism, and alter the rate of excretion.

PHARMACEUTICAL COMPOSITIONS

[0094] The pharmaceutical compositions of the present invention comprise an effective amount of a compound of the present invention formulated together with one or more pharmaceutically acceptable carriers. As used herein, the term "pharmaceutically acceptable carrier" means a non-toxic, inert solid or semi-solid filler, diluent, encapsulating material or auxiliary formulation of any kind. Some examples of materials that can serve as pharmaceutically acceptable carriers are sugars such as lactose, glucose and sucrose; starches, such as corn starch and potato starch; cellulose and its derivatives, such as sodium carboxymethylcellulose, ethylcellulose and cellulose acetate; powdered tragacanth; malt; gelatin; baby powder; excipients such as cocoa butter and suppository waxes; oils such as peanut oil, sunflower oil; safflower oil; Sesame oil; olive oil; corn oil and soybean oil; glycols such as propylene glycol; esters such as ethyl oleate and ethyl laurate; agar-agar; buffering agents such as magnesium hydroxide and aluminum hydroxide; alginic acid; pyrogen-free water; isotonic saline solution; Ringer's solution; Ethyl alcohol and phosphate buffer solutions, as well as other non-toxic compatible lubricants such as sodium lauryl sulfate and magnesium stearate, as well as coloring agents, release agents, coating agents, sweeteners, flavoring and odorizing agents, preservatives and antioxidants may also be present in the composition, at the judgment of the formulator. The pharmaceutical compositions of this invention can be administered to humans and other animals orally, rectally, parenterally, intracisternally, intravaginally, intraperitoneally, topically (as by powders, ointments, or drops), buccally, or as an oral or nasal spray.

[0095] The pharmaceutical compositions of the present invention can be administered orally, parenterally, by inhalation spray, topically, rectally, nasally, buccally, vaginally or via an implanted reservoir, preferably by oral administration or administration by injection. The pharmaceutical compositions of this invention can contain any conventional non-toxic pharmaceutically acceptable carriers, adjuvants or carriers. In some cases, the pH of the formulation can be adjusted with pharmaceutically acceptable acids, bases or buffers to improve the stability of the formulated compound or its delivery form. The term parenteral, as used herein, includes subcutaneous, intracutaneous, intravenous, intramuscular, intraarticular, intraarterial, intrasynovial, intrasternal, intrathecal, intralesional, and intracranial infusion or injection techniques.

[0096] Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups and elixirs. In addition to the active compounds, liquid dosage forms may contain inert diluents commonly used in the art such as, for example, water or other solvents, solubilizing and emulsifying agents, such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, dimethyl formamide, oils (in particular, cottonseed, peanut, corn, germ , olive, castor and sesame), glycerol, tetrahydrofurfuryl alcohol, polyethylene glycols and sorbitan fatty acid esters, and mixtures thereof. In addition to inert diluents, oral compositions can also include adjuvants, such as wetting agents, emulsifying and suspending agents, sweetening, flavoring and perfuming agents.

[0097] Injectable preparations, for example, sterile injectable aqueous or oleaginous suspensions, may be formulated according to the known technique using suitable dispersing or wetting agents and suspending agents. The sterile injectable preparation may also be a sterile injectable solution, suspension or emulsion in a parenterally acceptable non-toxic diluent or solvent, for example, as a solution in 1,3-butanediol. Among the acceptable vehicles and solvents that may be employed are water, Ringer's solution, U.S.P. and isotonic sodium chloride solution. In addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium. For this purpose, any bland fixed oil can be employed, including synthetic mono- or diglycerides. In addition, fatty acids such as oleic acid are used in the preparation of injectables.

[0098] Injectable formulations can be sterilized, for example, by filtration through a bacteria-retaining filter, or by incorporating sterilizing agents in the form of sterile solid compositions that can be dissolved or dispersed in sterile water or other sterile injectable media before use.

[0099] In order to prolong the effect of a drug, it is often desirable to delay the absorption of the drug from subcutaneous or intramuscular injection. This can be achieved by using a liquid suspension of crystalline or amorphous material with low water solubility. The rate of absorption of the drug then depends on its rate of dissolution which, in turn, may depend on crystal size and crystalline form. Alternatively, delayed absorption of a parenterally administered drug form is achieved by dissolving or suspending the compound in an oil vehicle. Injectable depot forms are made by forming microencapsulated matrices of the drug in biodegradable polymers such as polylactide-polyglycolide. Depending on the drug to polymer ratio and the nature of the polymer used, the rate of drug release can be controlled. Examples of other biodegradable polymers include poly(orthoesters) and poly(anhydrides). Depot injectable formulations are also prepared by entrapping the drug in liposomes or microemulsions that are compatible with body tissues.

[0100] Compositions for rectal or vaginal administration are preferably suppositories that can be prepared by mixing the compounds of this invention with suitable non-irritating excipients or carriers, such as cocoa butter, polyethylene glycol or a suppository wax, which are solid at room temperature but liquid at body temperature and therefore melt in the rectal or vaginal cavity and release the active compound.

[0101] Solid dosage forms for oral administration include capsules, tablets, pills, powders and granules. In such solid dosage forms, the active compound is mixed with at least one pharmaceutically acceptable, inert excipient or carrier such as sodium citrate or dicalcium phosphate and/or a) fillers or extenders such as starches, lactose, sucrose, glucose, mannitol, and silicic acid, b) binders such as, for example, carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose and acacia, c) humectants such as glycer d) agents such as agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates and sodium carbonate, e) solution delay agents such as paraffin, f) absorption accelerators, such as quaternary ammonium compounds, g) wetting agents such as, for example, cetyl alcohol and glycerol monostearate, h) absorbents such as kaolin and bentonite clay, and i) lubricants such as talc, stearate calcium act, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, and mixtures thereof. In the case of capsules, tablets and pills, the dosage form can also comprise buffering agents.

[0102] Solid compositions of a similar type can also be employed as fillers in soft and hard filled gelatin capsules using excipients such as lactose or milk sugar, as well as high molecular weight polyethylene glycols and the like.

[0103] The active compounds may also be in microencapsulated form with one or more excipients as noted above. Solid dosage forms of tablets, dragees, capsules, pills and granules can be prepared with coatings and shells such as enteric coatings, release control coatings and other coatings well known in the art of pharmaceutical formulation. In such solid dosage forms, the active compound can be mixed with at least one inert diluent, such as sucrose, lactose or starch. Such dosage forms may also comprise, as is normal practice, additional substances other than inert diluents, for example tabletting lubricants and other tabletting aids such as magnesium stearate and microcrystalline cellulose. In the case of capsules, tablets and pills, the dosage forms can also include buffering agents. They can optionally contain opacifying agents and can also be of a composition that they release the active ingredient(s) only, or preferentially, in a certain part of the intestinal tract, optionally, in a delayed manner. Examples of embedding compositions that can be used include waxes and polymeric substances.

[0104] Dosage forms for topical or transdermal administration of a compound of this invention include ointments, pastes, creams, lotions, gels, powders, solutions, sprays, inhalants or patches. The active component is mixed under sterile conditions with a pharmaceutically acceptable carrier and any necessary preservatives or buffers may be required. Ophthalmic formulations, eye drops, eye ointments, powders and solutions are also contemplated as being within the scope of the present invention.

[0105] Ointments, pastes, creams and gels may contain, in addition to an active compound of the drug product, excipients such as animal and vegetable fats, oils, waxes, paraffins, starch, tragacanth, cellulose derivatives, polyethylene glycols, silicones, bentonites, silicic acid, talc and zinc oxide, or mixtures thereof.

[0106] Powders and sprays may contain, in addition to the compounds of this invention, excipients such as lactose, talc, silicic acid, aluminum hydroxide, calcium silicates and polyamide powder, or mixtures of these substances. Sprays may also contain customary propellants, such as chlorofluorohydrocarbons.

[0107] Transdermal patches have the added advantage of providing controlled delivery of a compound to the body. Such dosage forms can be made by dissolving or dispensing the compound in the appropriate medium. Absorption enhancers can also be used to increase the flux of the compound across the skin. The rate can be controlled by providing a rate controlling membrane or by dispersing the compound in a polymeric matrix or gel.

[0108] Unless otherwise defined, all technical and scientific terms used in this document have the same meaning as commonly understood by those of common knowledge in the art. All publications, patents, disclosed patent applications and other references mentioned in this document are incorporated by reference in their entirety.

abbreviations

[0109] Abbreviations that have been used in the descriptions of the schemes and the examples below are: ACN for acetonitrile; BME for 2-mercaptoethanol; BOP for (benzotriazol-1-yloxy)tris(dimethylamino)phosphoniumhexafluorophosphate; BzCl for benzoyl chloride; CDI for carbonyldiimidazole; COD for cyclooctadiene; DABCO for 1,4-diazabicyclo[2.2.2]octane; DAST for diethylaminosulfide trifluoride; DABCYL for 6-(N-4'-carboxy-4-(dimethylamino)azobenzene)aminohexyl-1-O-(2-cyanoethyl)-(N,N-diisopropyl)-phosphoramidite; DBU for 1,8-Diazabicycloundec-7-ene; DCC for N,N'-dicyclohexylcarbodiimide; DCM for dichloromethane; DIAD for diisopropyl azodicarboxylate; DIBAL-H for diisobutylaluminum hydride; DIPEA for diisopropylethylamine; DMAP for N,N-dimethylaminopyridine; DME for ethylene glycol dimethyl ether; DMEM for Dulbecco's Modified Eagle's Medium; DMF for N,N-dimethylformamide; DMSO for dimethylsulfoxide; DSC for N,N'-disuccinimidyl carbonate; DPPA for diphenylphosphoryl azide; DUPHOS for EDANS for 5-(2-amino-ethylamino)-naphthalene-1-sulfonic acid; EDCI or EDC for 1-(3-diethylaminopropyl)-3-ethylcarbodiimide hydrochloride; EtOAc to ethyl acetate; EtOH for ethyl alcohol; HATU for O(7-Azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate; HCl for hydrochloric acid; Hoveyda Catalyst for Dichloro(o-isopropoxyphenylmethylene)(tricyclohexylphosphine)ruthenium(II); In for indium; KHMDS is potassium bis(trimethylsilyl) starch; Ms for mesil; NMM for N-4-methylmorpholine; NMI for N-methylimidazole; NMO for N-4-Methylmorpholine-N-Oxide; PyBrOP for bromo-tri-pyrolidino-phosphonium hexafluorophosphate; Ph for phenyl; RCM for ring closure metathesis; RT for reverse transcription; RT-PCR for reverse transcription polymerase chain reaction; TBME for tert-butyl methyl ether; TEA for triethylamine; Tf2O for trifluoromethanesulfonic anhydride; TFA for trifluoroacetic acid; THF for tetrahydrofuran; TLC for thin layer chromatography; (TMS)2NH for hexamethyldisilazane; TMSOTf for trimethylsilyl trifluoromethanesulfonate; TBS for t-Butyldimethylsilyl; TMS for trimethylsilyl; TPAP for tetrapropylammonium perruthenate; TPP or PPh3 for triphenylphosphine; TrCl for trityl chloride; DMTrCl for 4,4'-dimethoxytrityl chloride; tBOC or Boc for tert-butyloxycarbonyl.

Synthetic Methods

[0110] The compounds and processes of the present invention will be better understood in connection with the following schemes which illustrate the methods by which the compounds of the invention can be prepared, which are intended for illustration only and do not limit the scope of the invention. Various changes and modifications to the disclosed embodiments will be apparent to those skilled in the art and such changes and modifications, including, without limitation, those relating to the chemical structures, substituents, derivatives and/or methods of the invention can be made without departing from the spirit of the invention and the scope of the appended claims.

[0111] As illustrated in Scheme 1, novel bile acid analogues of the compound of formula (1-5) are prepared from the compound of formula (1-1), wherein R1, R7 are defined as above, P1 and P2 are hydroxyl protecting groups. Thus, the two hydroxyl groups of the compound of formula (1-1) are protected with groups P1 and P2 to provide the compound of formula (1-2). P1 and P2 can be the same or different. P1 and P2 can be any hydroxyl protecting group such as, but not limited to Ac, Bz, chloroacetyl, TES, TBS, MOM and Bn. A more detailed discussion of hydroxyl group protection procedures, reagents and conditions is described in the literature, for example by TW Greene and PGM Wuts in "Protective Groups in Organic Synthesis" 3rd ed., John Wiley & Son, Inc., 1999. Next, the compound of formula (1-2) is converted to the acyl azide compound of formula (1-3) using suitable reagent such as, but not limited to, DPPA. The reaction solvent can be, but not limited to, THF, DCM and toluene. The preferred solvent is THF. The reaction temperature is -20OC~40OC. Further rearrangement of the compound of formula (1-3) at elevated temperature and reaction with sulfonamide provides the compound of formula (1-4). Thereafter, deprotection of groups P1 and P2 provide the sulfonylurea compound of formula (1-5). A more detailed discussion of procedures, reagents and conditions for deprotection of the hydroxyl protecting group is described in the literature, for example by TW Greene and PGM Wuts in "Protective Groups in Organic Synthesis" 3rd ed., John Wiley & Son, Inc., 1999. Scheme 1

[0112] Scheme 2 illustrates the preparation of the urea compound of formula (2-2) from the compound of formula (1-3), wherein R10 , m, and R7 are defined as above, P1 and P2 are hydroxyl protecting groups. Thus, further rearrangement of the compound of formula (1-3) at elevated temperature and reaction with sulfonamide provides the compound of formula (2-1). Then, deprotection of groups P1 and P2 gives the urea compound of formula (2-2). A more detailed discussion of procedures, reagents and conditions for deprotection of the hydroxyl protecting group is described in the literature, for example by TW Greene and PGM Wuts in "Protective Groups in Organic Synthesis" 3rd ed., John Wiley & Son, Inc., 1999. Scheme 2

[0113] Scheme 3 illustrates the preparation of the sulfonamide compound of formula (3-4) from the compound of formula (1-3), wherein R1,m and R7 are defined as above, P1 and P2 are hydroxyl protecting groups. Thus, the compound of formula (1-3) is converted to the compound of formula (31) via the Curtius rearrangement. A more detailed discussion of the procedures, reagents and conditions for the Curtius rearrangement is described in the literature, for example by Jerry March in "Advanced Organic Chemistry" 4th ed., John Wiley & Son, Inc., 1992. Next, Boc deprotection of the compound of formula (3-1) in an acidic state gives amine compound of formula (3-2). Then, the compound of formula (3-2) is reacted with sulfonyl chloride to give the sulfonamide compound of formula (3-3). Further deprotection of the P1 and P2 hydroxyl protecting group to give the compound of formula (3-4). A more detailed discussion of procedures, reagents and conditions for deprotection of hydroxyl protecting groups and amino protecting groups is described in the literature, for example by TW Greene and PGM Wuts in "Protective Groups in Organic Synthesis" 3rd ed., John Wiley & Son, Inc., 1999. Scheme 3

[0114] An alternative procedure for preparing sulfonamide compounds of formula (4-3) is illustrated in Scheme 4, where R1, m and R7 are defined as above, P1 and P2 are hydroxyl protecting groups. The compound of formula (1 - 2) is coupled with sulfonamide using the proper coupling condition to give the compound of formula (4-1). The coupling reagent can be selected from, but not limited to, DCC, EDC, CDI, diisopropyl carbodiimide, BOP-Cl, PyBOP, PyAOP, TFFH and HATU. Suitable bases include, but are not limited to, triethylamine, diisopropylethylamine, DBU, N-methylmorpholine and DMAP. The coupling reaction is carried out in an aprotic solvent such as, but not limited to, CH2Cl2, DMF or THF. The reaction temperature can vary from 0oC to about 50oC. The compound of formula (4-1) is treated with reducing agent to give the compound of formula (4-2). The reducing agent can be selected but not limited to LiAlH4, LiBH4, DIBAL, BH3. The reaction is carried out in an aprotic solvent such as, but not limited to, CH2Cl2, DMF or THF. The reaction temperature can vary from 0oC to about 100oC. Further deprotection of the compound of formula (4-2) gives the compound of formula (4-3). A more detailed discussion of procedures, reagents and conditions for deprotection of the hydroxyl protecting group is described in the literature, for example by TW Greene and PGM Wuts in "Protective Groups in Organic Synthesis" 3rd ed., John Wiley & Son, Inc., 1999. Scheme 4

Examples

[0115] The compounds and processes of the present invention will be better understood in connection with the following examples, which are intended to be an illustration only and do not limit the scope of the invention. Various changes and modifications to the disclosed embodiments will be apparent to those skilled in the art, and these changes and modifications, including, without limitation, those relating to the chemical structures, substituents, derivatives, formulations, and/or methods of the invention can be made without departing from the spirit of the invention and the scope of the appended claims. Example 1: Step 1 - 1:

[0116] To a 1 dram vial were added 6(a)-ethyl-chenodeoxycholic acid (6-ECDCA) (100 mg, 0.24 mmol), anhydrous THF (3 mL), TBBSCl (107.5 mg, 0.71 mmol) and imidazole (57.2 mg, 0.84 mmol), respectively, and the reaction mixture was stirred at room temperature for 22 h. Transferred to a separatory funnel, diluted with EtOAC (50 mL) and washed with brine (10 mL, 1:1 v/v). It was dried, filtered and concentrated and the resulting white solid was dissolved in MeOH (10 mL) and K2CO3 (49.7 mg, 0.36 mmol) was added. The mixture was stirred at room temperature for 30 min. And it was cooled down to 0oC. Then the reaction mixture was acidified by adding 0.1 N HCl to pH <7 and diluted with EtOAc (30 mL). After washed with brine (10 mL), dried and concentrated, the resulting crude material was purified by CombiFlash (12 g SiO 2 , Acetone/Hexanes = 0 ~ 40%) to give compound (1) as a white solid, 88 mg, 65.6% yield over 2 steps. Step 1-2:

[0117] TBS-protected 6(a)-ethyl-cene-oxycholic acid compound (1) (125 mg, 0.23 mmol) was dissolved in THF (2.0 mL) and cooled to 0 °C. To the solution was added Et3N (64 µL, 0.46 mmol) and diphenylphosphoryl azide (74 µL, 0.35 mmol). The mixture was stirred at 0 °C for 1.5 h, quenched with brine and extracted with DCM (2x). The combined organic layer was dried over Na 2 SO 4 , filtered and concentrated in vacuo at 25°C. The residue was taken up in hexane, filtered through Na2SO4 and concentrated in vacuo at 25oC. The crude product compound (2) (150 mg) was used for the next step without purification. Step 1-3:

[0118] The acyl azide compound (2) obtained above (75 mg) was dissolved in toluene (2.5 ml) and refluxed for 5 hours. The mixture was cooled to room temperature and naphthalene-2-sulfonamide (58 mg, 0.24 mmol) and DBU (36 µL, 0.24 mmol) were added. The reaction mixture was stirred at room temperature for 1 hour, quenched with aq. 1 M HCl and extracted with EtOAc (2x). The combined organic layer was dried over Na2SO4, filtered and concentrated in vacuo. The residue was purified by SiO2 chromatography with 0-50% EtOAc/hexane as eluent to afford N-sulfonyl urea compound (3) (74 mg). Step 1-4:

[0119] The compound obtained above (3) (74 mg) was dissolved in MeOH (1.0 mL), followed by the addition of 1 drop of 37% conc. HCl. The mixture was stirred at room temperature for 10 minutes, quenched with sat. NaHCO3, and extracted with EtOAc (3x). The combined organic layer was dried over Na2SO4, filtered and concentrated in vacuo. The residue was chromatographed on SiO2 with 0-50% acetone/hexane as eluent to give the compound of Example 1 (44 mg).

[0120] Examples from Example 2 to Example 8 were prepared using the same procedure as used in Example 1. MS data and 1H NMR data are outlined in Table 9. Table 9 Example 8: Step 8-1:

[0121] 6(a)-ethyl-chenodeoxycholic acid (2.1 g, 2.38 mmol) was dissolved in toluene (11 ml). Formic acid (98%, 3.0 mL) and perchloric acid (70%, 20 μL) were added dropwise to the solution. The mixture was stirred at 105°C for 3.5 h and cooled to room temperature. The mixture was diluted with EtOAc and washed with brine. The organic layer was dried over Na2SO4, and concentrated in vacuo. Purification of the residue by SiO2 chromatography with 0-40% acetone/hexane compound provided (8 - 1) (730 mg). Step 8-2:

[0122] Compound (8-1) (730 mg, 1.53 mmol) was dissolved in THF (8.0 mL) and cooled to 0 °C. To the solution was added Et3N (425 µL, 3.06 mmol) and diphenylphosphoryl azide (347 µL, 1.61 mmol). The mixture was stirred at 0 °C for 1.5 h, quenched with water and extracted with EtOAc (2x). The combined organic layer was dried over Na 2 SO 4 , filtered and concentrated in vacuo at 25°C. The crude obtained above was dissolved in toluene (20 mL), stirred at 100 °C for 30 min and t-BuOH (1.5 mL) was added. The mixture was stirred at 100 °C for 18 h, cooled to room temperature, diluted with EtOAc and washed with water and brine. The organic layer was dried over Na2SO4, filtered and concentrated in vacuo. Purification of the residue by SiO2 chromatography using 0-20% EtOAc/hexane compound provided (8-2) (401 mg). LC/MS Found [M+NH4]+, 565.42. 1HNMR (500 MHz, CDCl3) 8.15 (1H, s) 8.04 (1H, s), 5.19 (1H, s), 4.71 (1H, brs), 4.41 (1H, brs), 3.19 (1H, brs), 3.03 (1H, brs), 1.44 (9H, s), 0, 95 (6H, brs), 0.90 (3H, t, J = 7.0 Hz), 0.65 (3H, s). Step 8-3:

[0123] Compound (8-2) (401 mg, 0.73 mmol) was dissolved in DCM (15 mL) and cooled to 0 °C. TFA (1.1 ml) was added dropwise and the reaction solution was warmed to room temperature and stirred for 1 h. The solvent was removed in vacuo. The residue was dissolved in DCM and washed with sat. NaHCO3. The organic layer was dried over Na2SO4, and concentrated in vacuo. Compound (8-3) (300 mg) was obtained as a white solid. LC/MS found [M+H]+, 448. Step 8-4:

[0124] The amine compound (8-3) (100 mg, 0.22 mmol) was dissolved in DCM (1.0 mL), followed by addition of Et3N (67 µL, 0.48 mmol) and phenylmethanesulfonyl chloride (46 mg, 0.24 mmol). The reaction mixture was stirred at room temperature for 18 hours, quenched with 5% NaHCO 3 and extracted with DCM (3x). The combined organic layers were dried over Na 2 SO 4 , filtered and concentrated in vacuo. Purification of the residue by SiO2 chromatography using 0-30% compound supplied EtOAc/hexane (8-4) (57 mg). LC/MS found [M+NH4]+, 619.38; [M+HCOOH-H]-, 646.27. 1HNMR (500 MHz, CDCl3) 8.15 (1H, s), 8.03 (1H, s), 7.38 (5H, s), 5.18 (1H, s), 4.70 (1H, br s), 4.24 (2H, s), 3.94 (1H, br s), 3.02 (1H, br s), 2.93 (1H, brs), 0.95 (3H, s), 0.89 (6H, m), 0.63 (3H, s). Step 8-5:

[0125] Compound (8-4) (57 mg, 0.095 mmol) was dissolved in MeOH (0.5 mL). To the solution was added aq. 50% NaOH solution (0.23 mL, 30 eq). The mixture was stirred at 50°C°C for 15 h, cooled to room temperature, quenched with 1M HCl and extracted with EtOAc (3x). The organic layer was dried over Na2SO4, and concentrated in vacuo. Purification of residue given compound of example 8. LC/MS found [M+HCOOH-H]-, 590.25. Example 9:

[0126] Compound (8-3) (57 mg, 0.095 mmol) was dissolved in MeOH (0.5 mL). To the solution was added aq. 50% NaOH solution (0.23 mL, 30 eq). The mixture was stirred at 50°C°C for 15 h, cooled to room temperature, quenched with 1M HCl and extracted with EtOAc (3x). The organic layer was dried over Na2SO4, and concentrated in vacuo. Purification of residue given compound of example 9. LC/MS found [M+1], 392.25. Example 10:

[0127] Acylsulfonamide (10-1) (100 mg, 0.18 mmol) was dissolved in THF (2 mL). To the solution at -78 °C added LiAlH 4 solution in THF (1.0 M in THF, 1.44 mL) dropwise. The mixture was warmed to room temperature and stirred for 1 h. The reaction was quenched at 0°C with saturated sodium potassium tartrate and stirred at room temperature for 14 hours. The mixture was extracted with EtOAc (2x) and the combined organic layer was dried over Na 2 SO 4 and concentrated in vacuo. Purification of the residue by SiO 2 chromatography with 0-50% EtOAc/hexane provided the compound of example 10 (42 mg). LC/MS found [M+HCOOH-H]-, 590.29. 1HNMR (500 MHz, CDCl3) 7.87 (2H, d, J = 7.5 Hz), 7.59 (1H, m), 7.52 (2H, m), 4.33 (1H, br s), 3.69 (1H, s), 3.41 (1H, br s), 3.92 (2H, br s), 0.90 (3H, t, J = 7.5 Hz), 0.89 (3H, s), 0.85 (3H, d, J = 5.5 Hz), 0.62 (3H, s). Example 11:

[0128] The compound of example 11 was prepared using a similar procedure to the compound of example 10. LC/MS found [M+HCOOH-H]-, 556.31. 1HNMR (500 MHz, CDCl3) 3.69 (1H, br s), 3.40 (1H, br s), 3.14 (1H, m), 3.09 (2H, m), 1.95 (1H, d, J = 12 Hz), 1.37 (6H, d, J = 5.5 Hz), 0.93-0.89 (9H, m), 0 .65 (3H, s).

[0129] Examples from Example 129 to Example 143 were prepared using a similar procedure as described above. MS data are outlined in Table 10. Table 10 Example 108:

[0130] 5-Phenylthiophene-2-sulfonamide (145 mg, 0.6 mmol) and DBU (91 mg, 0.6 mmol) in THF (5 mL) were added to a solution of compound (2a) (1 mL, 0.2 mmol) in toluene. The mixture was stirred at room temperature overnight. The mixture was quenched with water, extracted with ethyl acetate (50 mL), dried, filtered and concentrated. The residue was dissolved in MeOH (2 mL), then 1 drop of 37% HCl was added. The mixture was stirred at room temperature for 10 minutes, then diluted with ethyl acetate (50 mL) and washed sequentially with saturated sodium bicarbonate and brine. The organic layer was dried over Na2SO4, filtered and concentrated. The residue was purified by Flash-Prep-HPLC ((IntelFlash-1): Column, C18, Mobile Phase, MeCN/H2O, Detector, UV 254 nm) to give Example 108 (15.5 mg) as a white solid. Example 109: 1) . Compound Synthesis (109-1)

[0131] Compound (109-SM) (1g, 10mmol) was added to chlorosulfonic acid (10mL) at room temperature and the mixture was slowly heated to 100°C and then heated to 100-110°C for 2 hours. The reaction mixture was cooled, poured into 150 ml of crushed ice with stirring and extracted with ethyl acetate (20 ml*3). The organic layer was combined, washed with brine (30 ml), dried over Na 2 SO 4 , evaporated to give 1.25 g of the title compound (109 - 1) as a yellow oil which was used in the next step directly. two) . Compound Synthesis (109-2)

[0132] A solution of compound (109 - 1) (1.25 g, 6.48 mmol) in THF (40 mL) and ammonia (40 mL) was stirred at room temperature for 1.5 h. Then the solution was concentrated. The residue was purified by silica flash chromatography, gradient elution 40 to 100% EtOAc in petroleum ether to give the title compound of (109-2) (510mg, 45%) as a yellow solid. 3) . Summary of Example 109

[0133] Compound (109-2) (87.5 mg, 0.5 mmol) and DBU (76 mg, 0.5 mmol) in THF (5 mL) were added to a solution of compound (2a) (1 mL, 0.2 mmol) in toluene. The mixture was stirred at room temperature overnight. The mixture was quenched with water, extracted with ethyl acetate (50 mL), dried, filtered and concentrated. The residue was dissolved in MeOH (2 mL), then 1 drop of 37% HCl was added. The mixture was stirred at room temperature for 10 minutes, then diluted with ethyl acetate (50 mL) and washed sequentially with saturated sodium bicarbonate and brine. The organic layer was dried over Na2SO4, filtered and concentrated. The residue was purified by Flash-Prep-HPLC ((IntelFlash-1): Column, C18, Mobile Phase, MeCN/H2O, Detector, UV 220 nm) to give Example 109 (25.1 mg) as a white solid. Example 111: 1) . Compound Synthesis (111-1)

[0134] A solution of thiophene (6.0 g, 71.31 mmol) and 2-bromo-2-methylpropane (9.7 g, 70.79 mmol) in DCM (60 mL) was added dropwise to trichloroalumane (9.4 g, 70.50 mmol) in DCM (60 mL) at -78 °C. The resulting solution was stirred at -78°C for 2 h and warmed to room temperature overnight. The resulting mixture was diluted with DCM (200 mL) and washed sequentially with water (50 mL), 5% sodium hydroxide (50 mL) and brine (50 mL). The organic layer was dried over Na2SO4 and concentrated in vacuo to give 6.1 g (crude) of the desired compound as a yellow oil. two) . Compound Synthesis (111-2)

[0135] A solution of (111-1) (5 g, 35.7 mmol) in DCM (10 mL) was added dropwise to an ice-cold solution of chlorosulfonic acid (12.4 g, 107 mmol) in DCM (30 mL). The reaction mixture was stirred at 0°C for 30 min, then poured onto ice. The solution was extracted with DCM (20 mL*3). The combined organic layer was washed with H 2 O (30 mL) and saturated NaCl (30 mL), then dried over Na 2 SO 4 , filtered and evaporated to give 4.1 g (crude) of (111-2) as a yellow oil which was used in the next step directly. 3) . Compound Synthesis (111-3)

[0136] A solution of (111-2) (4.1 g, 17.2 mmol) in THF (40 mL) and ammonia (40 mL) was stirred at room temperature for 1.5 hours. So it was concentrated. The residue was purified by silica flash chromatography, gradient elution 40 to 100% EtOAc in petroleum ether to give the title compound (2.1, 56%) as a yellow solid. 4) . Summary of example 111

[0137] Compound (113-3) (110mg, 0.5mmol) in THF (5ml) was added to a solution of compound (2a) (1ml, 0.2mmol) in PhCH3. The mixture was stirred at room temperature overnight. The mixture was quenched with water, extracted with ethyl acetate (50 mL), dried, filtered and concentrated. The residue was dissolved in MeOH (2 mL), then 1 drop of 37% HCl was added. The mixture was stirred at room temperature for 10 minutes, then diluted with ethyl acetate (50 mL) and washed sequentially with saturated sodium bicarbonate and brine. The organic layer was dried over Na2SO4, filtered and concentrated. The residue was purified by Flash-Prep-HPLC ((IntelFlash-1): Column, C18, Mobile Phase, MeCN/H2O, Detector, UV 254 nm) to give Example 111 (29.4 mg) as a white solid. Example 112: 1) . Compound Synthesis (112-1)

[0138] A solution of Compound (112-SM) (4 mL) in DCM (10 mL) was added dropwise to an ice-cold solution of chlorosulfonic acid (10 mL) in DCM (30 mL). The reaction mixture was stirred for 2 h at room temperature and then poured onto ice. The solution was extracted with DCM (20 mL*3). The combined organic layer was washed with brine (30 mL), dried over Na 2 SO 4 , filtered and evaporated to give 1.1 g (crude) of (112-1) as a yellow oil, which was used in the next step directly. two) . Compound Synthesis (112-2)

[0139] A solution of compound (112-1) (1.1 g) in THF (40 mL) and ammonia (40 mL) was stirred at room temperature for 1.5 h. Then the solution was concentrated. The residue was purified by silica flash chromatography, gradient elution 50 to 100% EtOAc in petroleum ether to give 550 mg of the title compound (112-2) as a yellow solid. 3) . Summary of Example 112

[0140] Compound (112-2) (100 mg, 0.5 mmol) and DBU (76 mg, 0.5 mmol) in THF (5 mL) was added to a solution of compound (2a) (1 mL, 0.2 mmol) in toluene. The mixture was stirred at room temperature overnight. The mixture was quenched with water, extracted with ethyl acetate (50 mL), dried, filtered and concentrated. The residue was dissolved in MeOH (2 mL), then 1 drop of 37% HCl was added. The mixture was stirred at room temperature for 10 minutes, then diluted with ethyl acetate (50 mL) and washed sequentially with saturated sodium bicarbonate and brine. The organic layer was dried over Na2SO4, filtered and concentrated. The residue was purified by Flash-Prep-HPLC ((IntelFlash-1): Column, C18, Mobile Phase, MeCN/H2O, Detector, UV 254 nm) to give Example 112 (21.5 mg, 18%) as a white solid. Example 113: 1). Compound Synthesis (113-1)

[0141] In a 1000 mL round bottom flask purged and maintained with an inert nitrogen atmosphere, a solution of 6-ECCDCA (18.0 g, 0.04 mol, 1.00eq) in THF (200 mL), TEA (86.5 g, 0.86 mol, 20.0 eq), 4-dimethylaminopyridine (0.63 g, 0.004 mol, 0 .1eq), acetic anhydride (87.3 g, 0.86 mol, 20.0 eq). The resulting solution was stirred at 90°C for 15 hours. After cooling to room temperature, it was concentrated and the residue was dissolved in ethyl acetate (500 mL), then washed with water (100 mL*2), saturated NaCl (100 mL*2). The organic layer was dried over Na2SO4, filtered and concentrated. The residue was purified by silica flash chromatography, gradient elution 0 to 20% EtOAc in petroleum ether to give the desired compound (113 - 1) as a yellow solid (20.0 g, 92.6%). two). Compound Synthesis (113-2)

[0142] To a solution of (93-1) (20.0 g, 40 mmol) in TFA (150 mL) was added TFAA (63.0 g, 300 mmol). Then NaNO2 was added in 5 portions over 45 minutes at 0°C. After stirring at 0°C for 1 hour, the solution was moved to 40°C for 40 minutes. The solution was quenched with water after being cooled to room temperature, then extracted with ethyl acetate (200 mL * 3). The organic layer was dried over Na2SO4, filtered and concentrated. The residue was purified by silica flash chromatography, gradient elution 10 to 30% EtOAc in petroleum ether to give the desired compound as a yellow solid (12.2 g, 65.6%). 3). Compound Synthesis (113-3)

[0143] To a solution of (113-2) (11.7 g, 24.8 mmol) in CH3OH (100 mL) was added KOH (50.0 g, 892.8 mmol) and H2O (100 mL). The mixture was stirred at 90°C for 16 hours. The reaction mixture was quenched with 6N HCl to adjust the pH to 5 ~ 6, extracted with ethyl acetate (200 mL * 3). The organic layer was dried over Na2SO4, filtered and concentrated. The residue was purified by silica flash chromatography, gradient elution from 0 to 10% MeOH in DCM to give the desired compound as a yellow solid (9.0 g, 89%). 4). Compound Synthesis (113-4)

[0144] To a solution of (113-3) (5.0 g, 12.3 mmol) in THF (200 mL) was added imidazole (5.9 g, 86.1 mmol) and TBSCl (5.6 g, 36.9 mmol). The mixture was stirred at room temperature for 16 hours. The reaction mixture was quenched with 10% citric acid solution to adjust the pH to 5 ~ 6, extracted with ethyl acetate (150 mL *3). The organic layer was dried over Na2SO4, filtered and concentrated to give crude product (7.2 g, crude) as a yellow solid. To the yellow solid in CH3OH (250 mL) was added K2CO3 (2.3 g, 17.0 mmol). The mixture was stirred at room temperature for 4 hours. The reaction mixture was quenched with 10% citric acid solution to adjust the pH to 5 ~ 6, extracted with ethyl acetate (150 mL * 3). The organic layer was dried over Na2SO4, filtered and concentrated. The residue was purified by silica flash chromatography, gradient elution 10 to 20% EtOAc in petroleum ether to give the desired compound as a yellow solid (3.3 g, 56%). 5). Compound Synthesis (113-5)

[0145] To a solution of (113-4) (1.0 g, 2.0 mmol) in toluene (10 mL) was added TEA (4.2 mmol, 2.1 eq) and DPPA (2.1 mmol, 1.05 eq) sequentially at 0°C. The resulting mixture was stirred at 0°C for 1 hour. Then it was heated to 100°C and stirred for 5 hours. After cooling to room temperature, a 0.2M solution of compound (113-5) in toluene was obtained, which could be divided into several portions for the next reaction. 6). Summary of example 113

[0146] Cyclohexylbenzenesulfonamide (72 mg, 0.3 mmol) and DBU (0.153 g, 1 mmol) in THF (1 mL) were added to a solution of PH-ETA-C-005-5 (1 mL, 0.2 mmol) in toluene. The mixture was stirred at room temperature overnight. The mixture was quenched with water, extracted with ethyl acetate (20 mL * 3), dried over Na 2 SO 4 , filtered and concentrated. The residue was dissolved in MeOH (2 mL), then 1 drop of 37% HCl was added. The mixture was stirred at room temperature for 10 minutes. Then, the mixture was diluted with ethyl acetate (50 ml) and sequentially washed with saturated sodium bicarbonate and brine. The organic layer was dried over Na2SO4, filtered and concentrated. The residue was purified by Flash-Prep-HPLC ((IntelFlash-1): Column, C18, Mobile Phase, MeCN/H2O, Detector, UV 254 nm) to give Example 113 (44.7 mg) as a white solid. Example 114:

[0147] Compound (114-2) (71.7 mg, 0.3 mmol) and DBU (0.153 g, 1 mmol) in THF (1 mL) in a solution of (113-5) (1 mL, 0.2 mmol) in toluene. The mixture was stirred at room temperature overnight. The mixture was quenched with water, extracted with ethyl acetate (Example'114 dried over Na2SO4, filtered and concentrated. The residue was dissolved in MeOH (2 mL), then 1 drop of 37% HCl was added. The mixture was stirred at room temperature for 10 minutes, then diluted with ethyl acetate (50 mL) and washed sequentially with saturated sodium bicarbonate and brine. The organic layer was dried over Na2SO4, filtered and concentrated. residue was purified by Flash-Prep-HPLC ((IntelFlash-1): Column, C18, mobile phase, MeCN/H2O, Detector, UV 254 nm) to give Example 114 (12.7 mg) as a white solid. Example 115:

[0148] Compound (115-2) (63.9 mg, 0.3 mmol) and DBU (0.153 g, 1 mmol) in THF (1 mL) in a solution of (113-5) (1 mL, 0.2 mmol) in toluene. The mixture was stirred at room temperature overnight. The mixture was quenched with water, extracted with ethyl acetate (50 mL), dried over Na 2 SO 4 , filtered and concentrated. The residue was dissolved in MeOH (2 mL), then 1 drop of 37% HCl was added. The mixture was stirred at room temperature for 10 minutes. Then it was diluted with ethyl acetate (50 mL) and washed sequentially with saturated sodium bicarbonate and saline. The organic layer was dried over Na2SO4, filtered and concentrated. The residue was purified by Flash-Prep-HPLC ((IntelFlash-1): Column, C18; mobile phase, MeCN/H2O, Detector, UV 254 nm) to give example 115 as a white solid (25.9 mg). Example 116:

[0149] Compound (116-2) (72.3 mg, 0.3 mmol) and DBU (0.153 g, 1 mmol) in THF (1 mL) in a solution of (113-5) (1 mL, 0.2 mmol) in toluene. The mixture was stirred at room temperature overnight. The mixture was quenched with water, extracted with ethyl acetate (50 mL), dried over Na 2 SO 4 , filtered and concentrated. The residue was dissolved in MeOH (2 mL), then 1 drop of 37% HCl was added. The mixture was stirred at room temperature for 10 minutes. Then it was diluted with ethyl acetate (50 mL) and washed sequentially with saturated sodium bicarbonate and saline. The organic layer was dried over Na2SO4, filtered and concentrated. The residue was purified by Flash-Prep-HPLC ((IntelFlash-1): Column, C18, Mobile Phase, MeCN/H2O, Detector, UV 254 nm) to give Example 116 (24.2 mg) as a white solid. Example 117:

[0150] Compound (117-2) (60.3 mg, 0.3 mmol) and DBU (0.153 g, 1 mmol) in THF (1 mL) in a solution of (113-5) (1 mL, 0.2 mmol) in toluene. The mixture was stirred at room temperature overnight. The mixture was quenched with water, extracted with ethyl acetate (50 mL), dried over Na 2 SO 4 , filtered and concentrated. The residue was dissolved in MeOH (2 mL), then 1 drop of 37% HCl was added. The mixture was stirred at room temperature for 10 minutes. Then it was diluted with ethyl acetate (50 mL) and washed sequentially with saturated sodium bicarbonate and saline. The organic layer was dried over Na2SO4, filtered and concentrated. The residue was purified by Flash-Prep-HPLC ((IntelFlash-1): Column, C18, Mobile Phase, MeCN/H2O, Detector, UV 254 nm) to give Example 117 (32.8 mg) as a white solid. Example 118: Compound (118-2) (72 mg, 0.3 mmol) and DBU (0.153 g, 1 mmol) in THF (5 mL) in a solution of (113-5) (1 mL, 0.2 mmol) in toluene. The mixture was stirred at room temperature overnight. The mixture was quenched with water, extracted with ethyl acetate (20 mL * 3), dried over Na 2 SO 4 , filtered and concentrated. The residue was dissolved in MeOH (2 mL), then 1 drop of 37% HCl was added. The mixture was stirred at room temperature for 10 minutes, then diluted with ethyl acetate (50 mL) and washed sequentially with saturated sodium bicarbonate and brine. The organic layer was dried over Na2SO4, filtered and concentrated. The residue was purified by Flash-Prep-HPLC ((IntelFlash-1): Column, C18, Mobile Phase, MeCN/H2O, Detector, UV 254 nm) to give Example 118 (40 mg) as a white solid. Example 119: 1) Synthesis of compound (119-1)

[0151] A mixture of N,N-dimethylaniline (0.5 g, 4.13 mmol) and bistrimethylsilyl sulfate (0.5 g, 4.13 mmol) was heated at 160°C for 5 hours. The mixture was allowed to cool to room temperature and the resulting solid was isolated by filtration and washed with Et2O. The solid was then dissolved in H 2 O, and the solution concentrated in vacuo to give the title compound 600 mg (crude) as a white solid. two) . Compound Synthesis (119-2)

[0152] Compound (119-1) (600 mg) was added portionwise to a suspension of PCl5 (931 mg, 4.5 mmol) in DCM (20 mL) at 0°C. The mixture was then allowed to warm to room temperature and was then stirred at room temperature for 3 h. The mixture was concentrated in vacuo and the residue was dissolved in Et2O and H2O. The layers were separated and the organic layer was dried over Na 2 SO 4 , filtered and concentrated in vacuo to give the title compound 320 mg as a yellow solid, which was used directly without further purification. 3) . Compound Synthesis (119-3)

[0153] Ammonia (10 mL) was added to a solution of (119-2) (320 mg) in THF (10 mL) and stirred at room temperature for 1.5 h, then concentrated. The residue was purified by silica flash chromatography, gradient elution 40 to 100% EtOAc in petroleum ether to give the title compound (160mg, 54.7%) as a yellow solid. 4) . Summary of Example 119

[0154] Compound (119-3) (100 mg, 0.5 mmol) and DBU (76 mg, 0.5 mmol) in THF (5 ml) was added into a solution of compound (2a) (1 ml, 0.2 mmol) in toluene. The mixture was stirred at room temperature overnight. The mixture was quenched with water, extracted with ethyl acetate (50 mL), dried, filtered and concentrated. The residue was dissolved in MeOH (2 mL), then 1 drop of 37% HCl was added. The mixture was stirred at room temperature for 10 minutes, then diluted with ethyl acetate (50 mL) and washed sequentially with saturated sodium bicarbonate and brine. The organic layer was dried over Na2SO4, filtered and concentrated. The residue was purified by Flash-Prep-HPLC ((IntelFlash-1): Column, C18; mobile phase, MeCN/H2O, Detector, UV 220 nm) to give example 119 (24.3 mg) as a white solid. Example 121:

[0155] Compound (121-2) (72 mg, 0.3 mmol) and DBU (0.153 g, 1 mmol) in THF (1 mL) in a solution of (93-5) (1 mL, 0.2 mmol) in toluene. The mixture was stirred at room temperature overnight. The mixture was quenched with water, extracted with ethyl acetate (50 mL), dried over Na 2 SO 4 , filtered and concentrated. The residue was dissolved in MeOH (2 mL). Then, 1 drop of 37% HCl was added. The mixture was stirred at room temperature for 10 minutes. Then it was diluted with ethyl acetate (50 mL) and washed sequentially with saturated sodium bicarbonate and saline. The organic layer was dried over Na2SO4, filtered and concentrated. The residue was purified by Flash-Prep-HPLC ((IntelFlash-1): Column, C18, Mobile Phase, MeCN/H2O, Detector, UV 254 nm) to give Example 121 (29.7 mg) as a white solid. The following desired examples were prepared using procedures similar to those described above. MS data and 1H NMR data are outlined in Table 11. Table 11

ESSAY Human FXR Assay (NR1H4)

[0156] Determination of a ligand-mediated Gal4 promoter-driven transactivation to quantify ligand-binding mediated activation of FXR. FXR Reporter Assay Kit purchased from Indigo Bioscience (catalog number: IB00601) to determine the potency and effectiveness of compound developed by Enanta that can induce FXR activation. The primary application of this reporter assay system is to quantify the functional activity of human FXR. The assay utilizes non-human mammalian cells, CHO (Chinese Hamster Ovary) cells engineered to express human NR1H4 protein (referred to as FXR). The repeating cells also incorporate the cDNA encoding the beetle luciferase that catalyzes substrates and produces photon emissions. The luminescent intensity of the reaction is quantified using an Envision plate-reading luminometer. Reporter cells include the luciferase reporter gene functionally linked to an FXR-responsive promoter. Thus, quantifying changes in luciferase expression in treated reporter cells provides a sensitive surrogate measure of changes in FXR activity. EC50 and efficacy (normalized to CDCA set to 100%) is determined by XLFit. The assay is in accordance with the manufacturer's instructions. In summary, the assay was performed in white, 96-well plates, using the final volume of 100ul which contains cells with different doses of compounds. Retrieve Reporter Cells from -80°C storage. Perform a rapid thaw of frozen cells by transferring a 10 ml volume of 37°C cell recovery medium to the frozen cell tube. Recap the tube of Reporter Cells and immediately place it in a 37°C water bath for 5-10 minutes. Collect the Reporter Cell Suspension tube from the water bath. Disinfect the outer surface of the tube with a 70% alcohol swab and transfer to the cell culture hood. Dispense 90 μl of cell suspension into each well of the 96-well assay plate. Transfer the plate to a 37°C incubator, allowing the cells to adhere to the well. Dilute compounds in the Dilution Plate (DP) and administer the cells in the Assay Plate (AP). The DMSO content of the samples was maintained at 0.2%. Cells were incubated for an additional 22 hours before luciferase activities were measured. Thirty minutes before intending to quantify FXR activity, remove the Detection Substrate and Detection Buffer from the refrigerator and place it in a low-light area so that it can equilibrate to room temperature. Remove plate lid and discard all media contents by ejecting into an appropriate waste container. Gently lift the inverted plate onto a clean absorbent paper towel to remove residual droplets. The cells will remain firmly adhered to the bottoms of the wells. Add 100 µL of Luciferase Detection Reagent to each assay plate well. Let the assay plate sit at room temperature for at least 5 minutes after adding LDR. Set the instrument (Envision) to perform a single 5 second "plate shake" prior to reading the first assay well. Reading time can be 0.5 second (500mSec) per well. EC50 and efficacy (normalized to CDCA set to 100%) is determined by XLFit.

Human TGR5 (GPBAR1) Activity In Vitro Assay

[0157] The potency and efficacy of compounds of the invention at the TGR5 receptor were evaluated using in vitro assays performed using the DiscoverX Express Kit (CAMP Hunter™ eXpress GPBAR1 CHO-K1 GPCR Assay; Catalog Number: 95-0049E2CP2S) GPBAR1 (G-protein coupled bile acid receptor 1) encodes a member of the G protein-coupled receptor (GPCR) superfamily. Activation of GPBAR1 following ligand binding initiates a series of second message cascades that result in a cellular response. Treatment of CHO cells expressing GPBAR1 with bile acids induces intracellular cAMP production and receptor internalization. Compound potency and efficacy for GPBAR1 activation by measuring cyclic adenosine monophosphate (cyclic AMP or cAMP) levels in live cells using a competitive immunoassay based on Enzyme Fragment Complementation (EFC).

[0158] Next, after seeding the cells in the 96-well white microplate, place it in a humidified incubator at 37°C, 5% CO2 for 18-24 hours before testing. On the second day, proceed to the appropriate cAMP Hunter eXpress Protocol according to the manufacturer's instructions. Dissolve the agonist compound in DMSO at the desired stock concentration and prepare serial 3-fold dilutions of the agonist compound in Cell Assay Buffer. The concentration of each dilution should be prepared at 4X the final screening concentration (ie, 15 µL of compound + 45 µL of Cell Assay/cAMP Antibody Agent). For each dilution, the final solvent concentration must remain constant. Transfer 15 µL of diluted compound into the assay plate and incubate the plate for 30 minutes at 37°C. After the agonist incubation, add 60 µL of cAMP detection reagents/cAMP solution mix (cAMP Lysis Buffer, Substrate Reagent 1, cAMP Solution D) to the appropriate wells. Incubate for 1 hour at room temperature (23°C), protected from light. Add 60 μL of cAMP Solution A to the appropriate wells. Incubate for 3 hours at room temperature (23°C), protected from light. Read samples on the standard Envision luminescence plate reader. Calculation of average EC50 after log transformation.

[0159] To assess the FXR agonistic potency of the example compounds and the reference compounds, potency ranges were determined in the Human FXR Assay (NR1H4) as listed below in table 12. Efficacy was normalized to CDCA defined as 100%. (A=EC50 < 0.1 μM; B= 0.1 μM < EC50 < 1.0 μM; C=1.0 μM < EC50 < 10 μM; D= EC50 > 10 μM). Table 12

[0160] While this invention has been particularly shown and described with reference to preferred embodiments thereof, it will be understood by those skilled in the art that various changes in form and detail can be made therein, without deviating from the scope of the invention encompassed by the appended claims.

Claims (20)

1. Compound, characterized by the fact that it is represented by Formula I or a pharmaceutically acceptable salt thereof: wherein Ra is hydrogen or -C1-C8 alkyl; Rb is -C(O)NHSO2RI or -SO2R1; R1 is selected from the group consisting of: 1) Halogen; 2) Hydroxyl; 3) -C1-C8-alkyl; 4) Halo-C1-C4-alkyl; 5) Alkenyl -C2-C8; 6) -C2-C8-alkynyl; 7) -C3-C8-cycloalkyl; 8) Aryl; 9) Arylalkyl; 10) Alkylaryl; 11) Heterocycloalkyl; 12) Heteroaryl; 13) Heteroarylalkyl; 14) 15) 16) 17) 18) and 19) -NR10R11; R2 is hydrogen or -C1-C8-alkyl; m is selected from 0, 1, 2 and 3; R3 is hydrogen or hydroxyl; R4 is hydrogen or hydroxyl; R5 and R6 are each independently hydrogen or a hydroxyl protecting group; R7 is selected hydrogen or -C1-C8 alkyl; and R10 and R11 are each independently selected from hydrogen, -C1-C8 alkyl, -C2-C8 alkenyl, -C2-C8 alkynyl, -C3-C8 cycloalkyl, -C3-C8 heterocycloalkyl, or R10 and R11 are taken together with the nitrogen to which they are attached, forming a heterocyclic ring. 2. Compound, according to claim 1, characterized in that it is represented by Formula II or a pharmaceutically acceptable salt thereof: wherein Ra, Rb, R2, R3, R4, R7 and m are as defined in claim 1. 3. Compound, according to claim 1, characterized in that it is represented by Formula III or a pharmaceutically acceptable salt thereof: wherein Ra, Rb, R2, R3, R7 and m are as defined in claim 1. 4. Compound, according to claim 3, characterized in that it is represented by one of the formulas (III-1 to III-12) or a pharmaceutically acceptable salt thereof: wherein R1, R7, R10 and m are as defined in claim 3. 5. Compound, according to claim 1, characterized in that it is represented by Formulas IV-A and IV-B, or a pharmaceutically acceptable salt thereof: wherein R1 and m are as defined in claim 1. 6. Compound, according to claim 1, characterized in that it is selected from: (A) Compounds, according to Formula IV-A, in which R1 and are outlined for each compound below: and (B) Compounds, according to Formula IV-B, in which R1 and are delineated for each compound below: or a pharmaceutically acceptable salt thereof. 7. Compound, according to claim 1, characterized in that it is selected from: (A) Compounds, according to Formula VA, in which R1 and m are outlined for each compound below: and (B) Compounds, according to Formula VB, in which R1 and are delineated for each compound below: or a pharmaceutically acceptable salt thereof. 8. Compound, characterized by the fact that it is selected from the compounds listed below: or a pharmaceutically acceptable salt thereof. 9. Compound, according to claim 8, characterized in that it has the structure: or a pharmaceutically acceptable salt thereof. 10. Compound, according to claim 8, characterized in that it has the structure: or a pharmaceutically acceptable salt thereof. 11. Compound, according to claim 8, characterized in that it has the structure: or a pharmaceutically acceptable salt thereof. 12. Compound, according to claim 8, characterized in that it has the structure: or a pharmaceutically acceptable salt thereof. 13. Compound, according to claim 8, characterized in that it has the structure or a pharmaceutically acceptable salt thereof. 14. Compound, according to claim 8, characterized in that it has the structure: or a pharmaceutically acceptable salt thereof. 15. Use of a compound as defined in any one of claims 1 to 14, characterized in that it is in the manufacture of a medicament for the prevention or treatment of a disease or condition mediated by FXR. 16. Use according to claim 15, characterized in that the disease or condition mediated by FXR is selected from the group consisting of chronic liver disease, gastrointestinal disease, kidney disease, cardiovascular disease and metabolic disease. 17. Use according to claim 16, characterized in that the disease or condition mediated by FXR is a chronic liver disease selected from the group consisting of primary biliary cirrhosis (PBC), cerebrotendinous xanthomatosis (CTX), primary sclerosing cholangitis (CPS), drug-induced cholestasis, intrahepatic cholestasis of pregnancy, cholestasis associated with parenteral nutrition (PNAC), cholestasis associated with bacterial overgrowth or sepsis , autoimmune hepatitis, chronic viral hepatitis, alcoholic liver disease, non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), liver transplant-associated host-versus-graft disease, live-donor transplant liver regeneration, congenital hepatic fibrosis, choledocholithiasis, granulomatous liver disease, intra- or extrahepatic malignancy, Sjogren's syndrome, sarcoidosis, Wilson's disease, Gaucher's disease, hemochromatosis, and alpha deficiency 1-antitrypsin. 18. Use according to claim 17, characterized in that the disease or condition mediated by FXR is (a) a kidney disease selected from the group consisting of diabetic nephropathy, focal segmental glomerulosclerosis (FSGS), hypertensive nephrosclerosis, chronic glomerulonephritis, chronic transplant glomerulopathy, chronic interstitial nephritis and polycystic kidney disease; (b) a cardiovascular disease selected from the group consisting of atherosclerosis, arteriosclerosis, dyslipidemia, hypercholesterolemia and hypertriglyceridemia; or (c) a metabolic disease selected from the group consisting of insulin resistance, type I and type II diabetes and obesity. 19. Use according to claim 15, characterized in that the disease or condition mediated by FXR is non-alcoholic fatty liver disease, non-alcoholic steatohepatitis or primary biliary cirrhosis. 20. Pharmaceutical composition, characterized in that it comprises the compound as defined in any one of claims 1 to 14 and a pharmaceutically acceptable carrier.